AKT: Novus Biologicals

AF887

AKT [p Ser473] Antibody [Unconjugated] - Pan Specific

100 μg

AF887

120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (<a class=

120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (120-HD) for 15 minutes and MCF-7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (291-G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

<b>Biological Strategies Validation. </b>Akt phosphorylated at S473 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; <a class=

CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

F0110). To facilitate intracellular staining, cells were fixed with formaldehyde (FC004) and permeabilized with Methanol." class="big_lightbox">

Serum-starved MCF-7 human breast cancer cells were unstimulated (blue open histogram) or activated with 100 ng/mL Recombinant human IGF-1 (Catalog # 291-G1, dark orange filled histogram) for 15 minutes, then stained with Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887) followed by PE-conjugated Anti-Rabbit IgG Secondary Antibody (<a class=

F0110). To facilitate intracellular staining, cells were fixed with formaldehyde (FC004) and permeabilized with Methanol." class="big_thumb" />

291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox">

<b>Biological Strategies Validation. </b>Simple Western lane view shows lysates of MCF-7 human breast cancer cell line and A549 human lung carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (<a class=

291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

<b>Biological Strategies Validation. </b>Phospho-proteome profiling results of LX-2 cells co-cultured with C3A–CYP2E1 cells with or without DDC treatment and detection of the effect of DDC on intracellular kinases in LX-2 cells of co-cultures200 μg of total cell lysates from LX-2 cells co-cultured with C3A-2E1 cells with or without 100 μM DDC for 1 h were incubated with membranes of the human phospho-MAPK Array Kit according to the manufacturer's instructions. Phospho MAPK Array data were developed on X-ray films following exposure to chemiluminescent reagents. 20 μg aliquots of total cell lysates from LX-2 cells were subjected to Western blotting analysis. (A) Template showing the location of MAPK antibodies spotted onto the human phospho-MAPK Array Kit. (B) The activation status of ERK1/2 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (C) The activation status of p38 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (D) The activation status of Akt in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (E) The activation status of ERK1/2, p38 and Akt in LX-2 cells co-cultured with C3A cells after DDC treatment. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>DDC up-regulates MMP-1 through ERK1/2 and Akt activation(A) Co-cultures were treated with 100 μM DDC for the indicated time. Phosphorylation of ERK1/2, p38 and Akt were determined by Western blotting analysis. The corresponding non-phosphorylated ERK1/2, p38, Akt and beta -actin were used for protein loading control. (B) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of ERK1/2 inhibitor (U0126, 10 μM). MMP-1 protein levels were analysed by Western blotting. (C) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of p38 inhibitor (SB203580, 10 μM). MMP-1 protein levels were analysed by Western blotting. (D) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of Akt inhibitor (T3830, 50 μM). MMP-1 protein levels were analysed by Western blotting. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>Enhancement of autophagy by SLAMF5 is not mediated by inhibition of the mTORC1 complex. (A) The autophagy-inducer rapamycin was applied to control or SLAMF5-silenced cells for 4 h. Autophagy was monitored by western blot analysis using an LC3-specific antibody. A representative blot of four independent experiments is shown on the left, and densitometry of LC3-II to beta -actin ratios is depicted on the right panel. (B) Cells were treated as described in panel (A), except that they were cultured in the presence of 20 nM bafilomycin A1 (BafA1) for the last 2 h. Data were analyzed as described in panel (A). (C)SLAMF5-silenced or control monocyte-derived dendritic cells were activated with LPS/IFN gamma , and phosphorylation of Akt and p70S6K was analyzed with immunoblotting. Representative blots of four independent experiments are shown. The phospho-proteins were normalized to the total levels of Akt1 and p70S6K. Data are expressed as mean ± SD (**p < 0.01 and ***p < 0.001; ns, not significant). Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fimmu.2018.00062/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>p38 inhibitor SB203580 improves the up-regulation of MMP-1 by DDC through stimulating ERK1/2Co-cultures were treated with or without 100 μM DDC for 1 h in the presence or absence of p38 inhibitor (SB203580, 10 μM). Phosphorylation of ERK1/2, p38 and Akt in LX-2 cells of co-cultures were determined by Western blotting analysis. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effect of MON on the AKT/mTOR/FOXO3a signaling pathway in DEX-treated mice. The phosphorylation of mTOR, FOXO3a, and AKT was determined by Western blot (A). MON increased expression of phosphorylation of each protein in in gastrocnemius tissues of mice (B–D). Each value is the mean ± SD of three independent experiments. ## p < 0.01, ### p < 0.001 vs. Nor; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. DEX. Nor, normal group; DEX, dexamethasone-treated group; and MON, monotropein-administrated group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35565825), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effect of MON on the expression of p-Akt, p-FoxO3a, and p-mTOR in C2C12 myotubes. The phosphorylation of mTOR, FOXO3a, and AKT was determined by Western blot (A). MON increased expression of phosphorylation of each protein in C2C12 myotubes (B–D). Each value is the mean ± SD of three independent experiments. # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. Nor; * p < 0.05, *** p < 0.001 vs. DEX. Nor, normal group; DEX, dexamethasone-treated group; and MON, monotropein-administrated group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35565825), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF887U-100UG
Alexa Fluor 405	AF887V-100UG
Alexa Fluor 488	AF887G-100UG
Alexa Fluor 532	AF887X-100UG
Alexa Fluor 594	AF887T-100UG
Alexa Fluor 647	AF887R-100UG
Alexa Fluor 700	AF887N-100UG
Alexa Fluor 750	AF887S-100UG
Alexa Fluor Plus 405	AF887AFP405-100UG
Alexa Fluor Plus 488	AF887AFP488-100UG
Alexa Fluor Plus 555	AF887AFP555-100UG
Alexa Fluor Plus 594	AF887AFP594-100UG
Alexa Fluor Plus 647	AF887AFP647-100UG
Alexa Fluor Plus 680	AF887AFP680-100UG
Allophycocyanin	AF887A
Biotin	AF887B
CoraFluor 1	AF887CL1
DyLight 350	AF887D
DyLight 405	AF887E
DyLight 488	AF887K
DyLight 550	AF887L
DyLight 594	AF887M
DyLight 650	AF887W
DyLight 680	AF887Y
DyLight 755	AF887Z
HRP	AF887H
Janelia Fluor 525	AF887JF525
Janelia Fluor 549	AF887I
Janelia Fluor 646	AF887J
Janelia Fluor 669	AF887JF669
PerCP	AF887C
mFluor Violet 450 SE	AF887MFV450
mFluor Violet 500 SE	AF887MFV500
mFluor Violet 610 SE	AF887MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF887-SP

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MAB2055

AKT Antibody (281046) [Unconjugated] - Pan Specific

(1 Review)

(31 Publications)

100 μg

MAB2055

HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_lightbox">

Western blot shows lysates of MCF-7 human breast cancer cell line, MBA-MB-123 human breast cancer cell line, and TS1 mouse helper T cell line. PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # <a class=

HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." alt="Western blot shows lysates of MCF-7 human breast cancer cell line, MBA-MB-123 human breast cancer cell line, and TS1 mouse helper T cell line. PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_thumb" />

1775-KS), recombinant human Akt2, and recombinant human Akt3 (5 ng/lane). PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_lightbox">

Western blot shows Recombinant Human Active Akt1 (Catalog # <a class=

1775-KS), recombinant human Akt2, and recombinant human Akt3 (5 ng/lane). PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4." class="big_thumb" />

NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

Akt was detected in immersion fixed MDA-MB-231 human breast cancer cell line using Mouse Anti-Human/ Mouse/Rat Akt Monoclonal Antibody (Catalog # MAB2055) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # <a class=

NL007) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

MAB6815). Both antibodies were used at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

Akt pan specific (panels A and C) and Akt1 (E17K Mutation) (panels B and D) were detected in immersion fixed 293T human embryonic kidney cell line transfected with wild type (panels A and B) or E17K mutated (panels C and D) Akt1 using Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) and Mouse Anti-Human Akt1 (E17K Mutation) Monoclonal Antibody (Catalog # <a class=

MAB6815). Both antibodies were used at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox">

Akt was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (<a class=

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox">

Akt was detected in immersion fixed paraffin-embedded sections of mouse spleen tissue using Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (<a class=

VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_lightbox">

MCF-7 human breast cancer cell line was stained with Mouse Anti-Human/Mouse/Rat Akt Monoclonal Antibody (Catalog # MAB2055, filled histogram) or isotype control antibody (Catalog # <a class=

MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_thumb" />

Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Akt at approximately 62 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Activation of ErbB and WNT signaling pathways in kidneys from Cdh16Cre::Tfebfs mice.Transcriptional and biochemical analyses were performed on Cdh16Cre and Cdh16Cre::Tfebfs mice. (A,B) Tables show the relative increase of genes related to the ErbB (A) and WNT (B) pathways in the microarray analyses performed on kidneys from P0 Cdh16Cre::Tfebfs mice. Graphs show real-time PCR validations performed on kidneys from Cdh16Cre::Tfebfs mice at different stages (P0, P12, P30). Data are shown as the average (± SEM) of at least three Cdh16Cre::Tfebfs mice normalized versus wild-type mice. (C,D) Immunoblot analyses performed on (C) P30 kidney tissues and (D) primary kidney cells isolated from Cdh16Cre::Tfebfs mice to evaluate ErbB and WNT activation status. Each replicate is a distinct biological sample. ErbB signaling was assessed by looking at phosphoAKT (Ser473) to total AKT ratio, and phosphoERK1 (T202/Y204)/ERK2(T185/Y187) to total ERK ratio; WNT signaling was assessed by quantifying beta -catenin and CCND1 (Cyclin D1) protein levels. Graphs represent the densitometry quantification of Western blot bands. Values are normalized to actin when not specified and are shown as an average (± SEM) (*p<0.05, **p<0.01, ***p<0.001, two-sided, Student’s t test).DOI://dx.doi.org/10.7554/eLife.17047.00710.7554/eLife.17047.008Figure 3—source data 1.Complete list of 294 genes (represented by 361 probesets) significantly induced (FDR≤0.05) in the KSP_P0 microarray dataset (GSE62977).The genes are ranked by decreasing signed ratio (KSP_P0/CTL).DOI://dx.doi.org/10.7554/eLife.17047.00810.7554/eLife.17047.009Figure 3—source data 2.Complete list of 628 genes (represented by 729 probesets) significantly induced (FDR≤0.05) in the KSP_P14 microarray dataset (GSE63376).The genes are ranked by decreasing signed ratio (KSP_P14/CTL).DOI://dx.doi.org/10.7554/eLife.17047.009Complete list of 294 genes (represented by 361 probesets) significantly induced (FDR≤0.05) in the KSP_P0 microarray

Biochemical analysis of ErbB signaling.Immunoblot analysis performed on P90 kidneys from Cdh16Cre::Tfebfs mice (A) and P90 Cdh16CreErt2::Tfebfs animals induced with tamoxifen at P14 (B) and at P30 (C), respectively. Each replicate is a different biological sample. ErbB was analyzed by quantifying phosphoAKT (Ser473) to total AKT, and phosphoERK1 (T202/Y204)/ERK2(T185/Y187) to total ERK; graphs are the densitometry quantifications of Western blot bands normalized to wild-type line and are shown as an average (± SEM) (*p<0.05, **p<0.01, ***p<0.001, two-sided Student’s t test).DOI://dx.doi.org/10.7554/eLife.17047.011 Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/17047), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>ACPA inhibits p-Akt, induces p-JNK and affects levels of specific metabolites in NSCLC lines.a Principal component analysis (PCA) score plot: Metabolomics profiling of control and ACPA-treated A549, H1299, H358, and H838 cells. b Changes in variable importance in projection (VIP) values for 19 metabolites in A549 cells. c, d, e Changes in VIP values for 20 metabolites in H1299, H358, and H838 cells. Significantly changed metabolites (*p < 0.05, indicated by arrows) were matched to apoptotic pathways. f, g, h, i Increase and decrease in several metabolites of ACPA-treated A549, H1299, H358, and H838 cells (*p < 0.05). j Simple Western showing total Akt, p-Akt (S473), total JNK46 and JNK54 and p-JNK46 and p-JNK54 (T183/Y185) in A549 cells at 24 hours after treatment with IC50 dose of ACPA. k Relative expression levels of Akt and p-Akt for control and ACPA-treated A549 cells after normalization by total vinculin protein. l Relative expression levels of JNK (46 and 54 kDa) and p-JNK for control and ACPA-treated A549 cells after normalization by total vinculin protein. *p < 0.05, Student’s t-test. All tests were done in quadruplicates. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33431819), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , ,

Applications: , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	IC2055U-100UG
Alexa Fluor 405	IC2055V-100UG
Alexa Fluor 488	IC2055G-100UG
Alexa Fluor 532	FAB2055X
Alexa Fluor 594	IC2055T-100UG
Alexa Fluor 647	IC2055R-100UG
Alexa Fluor 700	IC2055N-100UG
Alexa Fluor 750	IC2055S-100UG
Alexa Fluor Plus 405	FAB2055AFP405-100UG
Alexa Fluor Plus 488	FAB2055AFP488-100UG
Alexa Fluor Plus 555	FAB2055AFP555-100UG
Alexa Fluor Plus 594	FAB2055AFP594-100UG
Alexa Fluor Plus 647	FAB2055AFP647-100UG
Alexa Fluor Plus 680	FAB2055AFP680-100UG
Allophycocyanin	IC2055A
Biotin	FAB2055B
CoraFluor 1	FAB2055CL1
DyLight 350	FAB2055D
DyLight 405	FAB2055E
DyLight 488	FAB2055K
DyLight 550	FAB2055L
DyLight 594	FAB2055M
DyLight 650	FAB2055W
DyLight 680	FAB2055Y
DyLight 755	FAB2055Z
FITC	FAB2055F
HRP	FAB2055H
Janelia Fluor 525	FAB2055JF525
Janelia Fluor 549	FAB2055I
Janelia Fluor 646	FAB2055J
Janelia Fluor 669	FAB2055JF669
PerCP	FAB2055C
Phycoerythrin	IC2055P
mFluor Violet 450 SE	FAB2055MFV450
mFluor Violet 500 SE	FAB2055MFV500
mFluor Violet 610 SE	FAB2055MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC2055A
Carrier Free	IC2055P
Carrier Free	MAB2055-SP
Carrier Free	IC2055V-100UG
Carrier Free	IC2055U-100UG
Carrier Free	IC2055T-100UG
Carrier Free	IC2055S-100UG
Carrier Free	IC2055R-100UG
Carrier Free	IC2055N-100UG
Carrier Free	IC2055G-100UG

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(select up to 3 products from this page to compare)

NB100-56749

AKT1 [p Ser473] Antibody (104A282) - BSA Free

(2 Reviews)

(15 Publications)

0.1 mg

NB100-56749

Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - IHC analysis of an FFPE human breast carcinoma tissue section using 1:250 dilution of phospho Ser473 AKT1 antibody (clone 104A282) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10 mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of phosphor-AKT (Ser-473) in the cancer cells as well as the stromal cells. Staining was performed by Histowiz.

Immunohistochemistry-Frozen: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - Rat mammary carcinoma tissue section stained with AKT1 [p Ser473] Antibody (104A282). IHC-Fr image submitted by a verified customer review.

SP1/PTEN mediates lung infection in mice and the activity of the Akt signaling pathway. A, survival days of the mice after si-SP1 and si-PTEN administration (p < 0.01, the Kaplan-Meier analysis); B, bacterial load in murine lung tissues examined by CFU analysis (*p < 0.05, the one-way ANOVA); C, infiltration of inflammatory cells in murine lung tissues examined by HE staining (*p < 0.05, the one-way ANOVA); D, epithelial cell apoptosis in the murine lung tissues after Mtb infection examined by the TUNEL assay (*p < 0.05, the one-way ANOVA); E, fibrosis in murine lung tissues determined by Masson’s trichrome staining (*p < 0.05, the one-way ANOVA); F-G, protein levels of SP1 and PTEN and the Akt phosphorylation in murine lung tissues determined by western blot analysis (*p < 0.05, the one-way ANOVA). For animal studies, n = 6 in each group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35420971), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EZH2/HMGA2 regulates the PI3K/AKT signaling. MI rats were subjected to injection of EZH2-OE or EZH2-NC (MI + MSC-EXO + EZH2 NC/EZH2 OE) and EZH2-OE + HMGA2-OE or EZH2-OE + HMGA2-NC. A HMGA2 protein expression in myocardial tissues in response to EZH2-OE + HMGA2-OE examined using western blot (Additional file 5: Fig. S5). B Weight change in rats after EZH2-OE + HMGA2-OE treatment. C Changes in HMI in rats after EZH2-OE + HMGA2-OE treatment. D Measurement of Dd, Sd, and LVEF by echocardiography at 2 weeks after LAD in rats. E Measurement of LVMI by echocardiography in rats. F KEGG pathway analysis of gene enrichment pathways [29]. G Changes in PI3K/AKT pathway in rat myocardium examined using western blot (Additional file 6: Fig. S6). All data are expressed as mean +/- SD (n = 6/group, *p < 0.05 vs. Sham, DMSO, MI + MSC-EXO + EZH2-NC or EZH2-OE + HMGA2-NC group determined by unpaired t test or one-way ANOVA) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35264108), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Rb, Am, Ch, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Rb, Am, Ch, Ze

Applications:

,

Applications: ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-187A

Alt Catalog# IMG-187A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-56749AF350
Alexa Fluor 405	NB100-56749AF405
Alexa Fluor 488	NB100-56749AF488
Alexa Fluor 532	NB100-56749AF532
Alexa Fluor 594	NB100-56749AF594
Alexa Fluor 647	NB100-56749AF647
Alexa Fluor 700	NB100-56749AF700
Alexa Fluor 750	NB100-56749AF750
Alexa Fluor Plus 405	NB100-56749AFP405
Alexa Fluor Plus 488	NB100-56749AFP488
Alexa Fluor Plus 555	NB100-56749AFP555
Alexa Fluor Plus 594	NB100-56749AFP594
Alexa Fluor Plus 647	NB100-56749AFP647
Alexa Fluor Plus 680	NB100-56749AFP680
Biotin	NBP2-27399B
DyLight 350	NB100-56749UV
DyLight 405	NB100-56749V
DyLight 488	NB100-56749G
DyLight 550	NB100-56749R
DyLight 594	NB100-56749DL594
DyLight 650	NB100-56749C
DyLight 680	NB100-56749FR
DyLight 755	NB100-56749IR
FITC	NBP2-27399F
HRP	NB100-56749H
Janelia Fluor 549	NB100-56749JF549
Janelia Fluor 646	NB100-56749JF646

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-27399
Azide and BSA Free	NBP2-27399SS
BSA Free	NB100-56749SS

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MAB887

AKT [p Ser473] Antibody (545007) [Unconjugated] - Pan Specific

(12 Publications)

100 μg

MAB887

222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF-7 human breast cancer cell line, and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 nm Calyculin for 30 minutes, 1 µg/mL insulin for 5 minutes, or 100 ng/mL Recombinant Human PDGF-AB (Catalog # <a class=

222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF-7 human breast cancer cell line, and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 nm Calyculin for 30 minutes, 1 µg/mL insulin for 5 minutes, or 100 ng/mL Recombinant Human PDGF-AB (Catalog # 222-AB) for 20 minutes. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

291-G1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox">

<b>Biological Strategies Validation. </b>Simple Western lane view shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (Catalog # <a class=

291-G1) for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 65 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse Phospho-Akt (S473) Pan Specific Monoclonal Antibody (Catalog # MAB887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

PTEN status in tested models (A) PTEN and pAKT levels in kidney cancer cell lines, IOD calculated relative to Vinculin (B) Relative to GusB expression of Pten in RNA isolates from kidney cancer and healthy kidney tissues (C) graphical workflow. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35625614), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

,

Applications: ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB887U
Alexa Fluor 405	FAB887V
Alexa Fluor 488	IC7794G
Alexa Fluor 532	FAB887X
Alexa Fluor 594	FAB887T
Alexa Fluor 647	FAB887R
Alexa Fluor 700	FAB887N
Alexa Fluor 750	FAB887S
Allophycocyanin	FAB887A
Biotin	FAB887B
CoraFluor 1	FAB887CL1
DyLight 350	FAB887D
DyLight 405	FAB887E
DyLight 488	FAB887K
DyLight 550	FAB887L
DyLight 594	FAB887M
DyLight 650	FAB887W
DyLight 680	FAB887Y
DyLight 755	FAB887Z
FITC	FAB887F
HRP	FAB887H
Janelia Fluor 525	FAB887JF525
Janelia Fluor 549	FAB887I
Janelia Fluor 646	FAB887J
Janelia Fluor 669	FAB887JF669
PerCP	FAB887C
Phycoerythrin	IC7794P
mFluor Violet 450 SE	FAB887MFV450
mFluor Violet 500 SE	FAB887MFV500
mFluor Violet 610 SE	FAB887MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC7794G
Carrier Free	IC7794P
Carrier Free	MAB887-SP

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NBP2-01724

AKT1 Antibody (OTI4D6)

(1 Review)

(2 Publications)

0.1 ml

NBP2-01724

Immunocytochemistry/Immunofluorescence: AKT1 Antibody (OTI4D6) [NBP2-01724] - ICC/IF of COS7 cells transiently transfected by pCMV6-ENTRY AKT1.

Western Blot: AKT1 Antibody (OTI4D6) [NBP2-01724] - WB analysis of extracts (10 ug) from a mouse cell line and 3 different mouse tissues using AKT1 antibody at a 1:200 dilution.

Immunohistochemistry-Paraffin: AKT1 Antibody (OTI4D6) [NBP2-01724] - IHC-P staining of paraffin-embedded human prostate tissue using anti-AKT1 antibody. Heat-induced epitope retrieval in 10 mM citric buffer, pH 6.0, 100C for 10 min.

Flow Cytometry: AKT1 Antibody (OTI4D6) [NBP2-01724] - Flow cytometric analysis of Jurkat cells using anti-AKT1 antibody (red), compared to a nonspecific negative control antibody (blue).

Western Blot: AKT1 Antibody (OTI4D6) [NBP2-01724] - HEK293T cells were transfected with the pCMV6-ENTRY control (left lane) or pCMV6-ENTRY AKT1 (right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-AKT1.

Immunohistochemistry-Paraffin: AKT1 Antibody (OTI4D6) [NBP2-01724] - IHC-P staining of paraffin-embedded adenocarcinoma of human endometrium tissue using anti-AKT1 antibody. Heat-induced epitope retrieval in10 mM citric buffer, pH 6.0, 100C for 10 min.

Immunohistochemistry-Paraffin: AKT1 Antibody (OTI4D6) [NBP2-01724] - IHC-P staining of paraffin-embedded carcinoma of human bladder tissue using anti-AKT1 antibody. Heat-induced epitope retrieval in 10 mM citric buffer, pH 6.0, 100C for 10 min.

Immunohistochemistry-Paraffin: AKT1 Antibody (OTI4D6) [NBP2-01724] - IHC-P staining of paraffin-embedded carcinoma of human kidney tissue using AKT1 antibody. Heat-induced epitope retrieval in 10 mM citric buffer, pH 6.0, 100C for 10 min.

Immunohistochemistry-Paraffin: AKT1 Antibody (OTI4D6) [NBP2-01724] - IHC-P staining of paraffin-embedded human kidney tissue using anti-AKT1 antibody. Heat-induced epitope retrieval in 10 mM citric buffer, pH 6.0, 100C for 10 min.

Reactivity:

Hu, Mu, Rt, Ca, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Pm

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-70135

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NBP1-51602

AKT1 Antibody (3A3)

(3 Reviews)

(3 Publications)

0.1 ml

NBP1-51602

Western Blot: AKT1 Antibody (3A3) [NBP1-51602] - Analysis of extracts from COS cells using AKT1 antibody at 1:1000. WB image submitted by a verified customer review.

Immunohistochemistry-Paraffin: AKT1 Antibody (3A3) [NBP1-51602] - Analysis of human endometrial cancer using AKT1 antibody at 1:25. IHC-P image submitted by a verified customer review.

Reactivity:

Hu, Mu, Pm

Species Glossary

Reactivity: Hu, Mu, Pm

Applications:

, , ,

Applications: , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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MAB7419

AKT1 [p Thr308] Antibody (658320) [Unconjugated]

(7 Publications)

100 μg

MAB7419

220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF-BB (Catalog # <a class=

220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of Jurkat human acute T cell leukemia cell line and NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human PDGF-BB (Catalog # 220-BB) for 20 minutes or 100nM Calyculin A for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-Akt1 (T308) Monoclonal Antibody (Catalog # MAB7419) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-Akt1 (T308) at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

220-BB) using Mouse Anti-Human Phospho-Akt1 (T308) Mono-clonal Antibody (Catalog # MAB7419) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox">

<b>Biological Strategies Validation. </b>Akt1 phosphorylated at T308 was detected in immersion fixed CCD-1070Sk human foreskin fibroblast cell line stimulated with Recombinant Human PDGF-BB (Catalog # <a class=

220-BB) using Mouse Anti-Human Phospho-Akt1 (T308) Mono-clonal Antibody (Catalog # MAB7419) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

<b>Biological Strategies Validation. </b>ASPP2 blocks Gal-1 dependent nanoclustering and halts oncogenic H-ras induced transformation.Nanoclustering-FRET analysis in HEK cells coexpressing mGFP- and mCherry-tagged (A, B) H-rasG12V or (C, D) K-rasG12V. Cells were analysed after overexpression of either Gal-1 or ASPP2 plasmids, or both (1:1 ratio). Plotted are the means ± SEM, n = 3. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left) or K-rasG12V (right) alone or together with Gal-1 and ASPP2. Statistical significance of differences was examined using t-test (n = 3; *, p<0.05, **, p< 0.01, ***, p<0.001). (F, G) Colony survival assay of NIH/3T3 cells stably expressing (F) H-rasG12V or (G) K-rasG12V and transiently expressing indicated constructs. Colony survival was graphed based on mean foci areas calculated from at least 4 independent biological repeats. (A-D, F-G) Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (ns, not significant; *, p<0.05; **, p<0.01; ****, p<0.0001). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.

<b>Biological Strategies Validation. </b>ASPP2 increases oncogenic H-ras, K-ras and N-ras-effector-recruitment, as well as ERK- and AKT-signalling.(A) Left, scheme explaining effector-recruitment FRET analysis in HEK cells. Right, examples of FLIM-FRET images of HEK cells from the different FRET samples as indicated. (B-D) Effector-recruitment FRET analysis in HEK cells coexpressing (B) mGFP-H-rasG12V, (C) mGFP-K-rasG12V or (D) mGFP-NrasG12V and mRFP-RBD from C-Raf. The effect of Gal-1 or ASPP2 expression on effector-recruitment FRET was compared to control samples. (E) Representative Western blots from HEK cells expressing mGFP-H-rasG12V (left), K-rasG12V (middle) or N-rasG12V (right) without or with Gal-1 or ASPP2. Statistical significance of differences between controls and treated samples was examined using one-way ANOVA (mean ± SEM, n = 3; ns, not significant; *, p<0.05, **, p< 0.01, ***, p<0.001, ****, p< 0.0001). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0159677), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

,

Applications: ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB7419U-100UG
Alexa Fluor 405	FAB7419V-100UG
Alexa Fluor 488	FAB7419G-100UG
Alexa Fluor 532	FAB7419X-100UG
Alexa Fluor 594	FAB7419T-100UG
Alexa Fluor 647	FAB7419R-100UG
Alexa Fluor 700	FAB7419N-100UG
Alexa Fluor 750	FAB7419S-100UG
Alexa Fluor Plus 405	FAB7419AFP405-100UG
Alexa Fluor Plus 488	FAB7419AFP488-100UG
Alexa Fluor Plus 555	FAB7419AFP555-100UG
Alexa Fluor Plus 594	FAB7419AFP594-100UG
Alexa Fluor Plus 647	FAB7419AFP647-100UG
Alexa Fluor Plus 680	FAB7419AFP680-100UG
Allophycocyanin	FAB7419A
Biotin	FAB7419B
CoraFluor 1	FAB7419CL1
DyLight 350	FAB7419D
DyLight 405	FAB7419E
DyLight 488	FAB7419K
DyLight 550	FAB7419L
DyLight 594	FAB7419M
DyLight 650	FAB7419W
DyLight 680	FAB7419Y
DyLight 755	FAB7419Z
FITC	FAB7419F
HRP	FAB7419H
Janelia Fluor 525	FAB7419JF525
Janelia Fluor 549	FAB7419I
Janelia Fluor 646	FAB7419J
Janelia Fluor 669	FAB7419JF669
PE	FAB7419P
PerCP	FAB7419C
mFluor Violet 450 SE	FAB7419MFV450
mFluor Violet 500 SE	FAB7419MFV500
mFluor Violet 610 SE	FAB7419MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB7419-SP

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NBP1-69923

AKT1 [p Ser473] Antibody (17F6.B11) - BSA Free

(2 Publications)

0.1 mg

NBP1-69923

Western Blot: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Western Blot of AKT1 [p Ser473] antibody (17F6.B11). A: Lane 1) PDGF stimulated NIH 3T3 cells [10ul]. Lane 2) NIH 3T3 cells [10ul]. Lane 3) Hela whole cell lysate [10ul] (weak signal). B: Lane 4) GST negative control protein [100ng]. Lane 5) GST negative control protein [25ng]. Lane 6) AKT 1 recombinant protein [100ng]. Lane 7) AKT 1 recombinant protein [25ng].Block: 5% BSA overnight at 4C.Primary antibody: Monoclonal anti-AKT antibody used at 1:1000 for overnight at 4C.Secondary antibody: HRP Conjugated goat anti-mouse 1:25K for 45 min at RT.Detection: TMB for 20 minutes, rinsed with deionized water, dried and scanned on conventional flatbed scanner.

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - High resolution STED immunofluorescence nanoscopy of AKT11 [p Ser473] antibody (17F6.B11). Tissue: A431 cells. The merge images (A) and at high magnification (B) show phosphorylated AKT1 colocalized with the distal microtubules. Fixation: 4% paraformaldehyde for 5 min and after washes blocked with 10% NGS/0.2% Triton X-100 for 30 min.Antigen retrieval: serum deprivation for 12 h.Primary antibody: AKT1 pS473 antibody at 10 ug/mL and alpha-tubulin (cyan) at 1.4 ug/mL for 1 h at RT.Secondary antibody: Atto 647N anti-Mouse IgG (ATTO TEC GmbH), and DyLight(TM)488 anti-Rabbit IgG were used at 1.0 ug/mL for 1h at RT for indirect detection.Localization: AKT1 pS473 is in the cytoplasm and also organized at the periphery of the cell.Staining: AKT1 pS473 as red signal with bis-benzimide (blue) nuclear counterstain.

Immunohistochemistry: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Immunohistochemistry of AKT11 [p Ser473] antibody (17F6.B11). Tissue: human prostate tissue. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 20 ug/mL for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.Staining: AKT1 pS473 as precipitated red signal with hematoxylin purple nuclear counterstain.

AKT1-p-Ser473-Antibody-17F6-B11-Western-Blot-NBP1-69923-img0037.jpg

<b>Biological Strategies Validation. </b>Western Blot: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Western Blot of AKT11 [p Ser473] antibody (17F6.B11). Lane 1: untreated NIH/3T3 cell lysates. Lane 2: detects phosphorylated AKT1 (indicated by arrowhead at ~56 kDa) on PDGF stimulated NIH/3T3 cell lysates. Load: 10 ug per lane.Primary antibody: AKT1 pS473 antibody at 1:10,000 in TBS with 0.05% Tween-20 with 1% BSA, for 1 h at 4 C.Secondary antibody: HRP conjugated Gt-a-Mouse IgG was used at a 1:20,000 dilution for 1 h at 4 C with FemtoMax(TM) enhanced chemiluminescent reagent.

Western Blot: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Western Blot of AKT11 [p Ser473] antibody (17F6.B11). Lane 1: A431 cells. Lane 2: A431 cells stimulated for 15 min with EGF. Load: 35 ug per lane.Primary antibody: AKT1pS473 antibody at 1:400 for overnight at 4C.Secondary antibody: DyLight(TM)649 Conjugated Anti-AKT1 pS473 Monoclonal Antibody at 1:10,000 for 45 min at RT.Block: Blocking Buffer for Fluorescent Western Blotting overnight at 4C.Predicted/Observed size: 56kDa. Other band(s): none.

Western Blot: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Western Blot of AKT1 [p Ser473] antibody (17F6.B11). Lane 1: unstimulated NIH/3T3 lysates contain inactive unphosphorylated AKT11, green band. Lane 2: PDGF stimulated NIH/3T3 lysate contains both inactive (green band) and activated phosphorylated AKT11 (red band). Load: 10 ug per lane.Primary antibody: rabbit anti-AKT1 (pan) and mouse anti-AKT1 pS473 specific antibodies at 1:400 for overnight at 4C.Secondary antibody: DyLight(TM) 549 conjugated anti-rabbit IgG (green) and DyLight(TM) 649 conjugated anti-mouse IgG (red) secondary antibodies at 1:10,000 for 45 min at RT.Block: 5% BLOTTO overnight at 4C.

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Analysis of DyLight 488 conjugate of NBP1-69923. This image shows anti-histone detection using a DyLight 488 conjugate (green). Anti-Tubulin was detected using a DyLight 549 conjugate (red). Nuclei were counter-stained using DAPI (blue).

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923]

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Immunofluorescence Microscopy of AKT11 [p Ser473] antibody (17F6.B11) using STED nanoscopy to evaluate AKT1 activation and migration. Tissue: A431 cells. Antigen retrieval: Panel A: serum starved, unstimulated cells. Panel B: serum starved, EGF stimulated for 15 mins.A massive increase in AKT1-pS473 activation, as measured by intensity signal, peaked at 15 minutes and was associated with depolymerized tubulin.Staining: Panel A shows STED data (AKT1-pS473, red channel) collected simultaneously with confocal signal (a-tubulin, green channel). Upon stimulation of cells with EGF, a rapid activation of AKT1 is observed (Panel B) along with a coincident change in the tubulin organization (yellow signal), as well as an extensive cell shape-change (cell membrane folding) and accumulation of AKT1pS473 at the cell periphery.

Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Analysis of Biotin conjugate of NBP1-69923. 20 ug/mL for 1 h at RT Secondary antibody: Streptavidin Conj. HRP 10 ug/ml Localization: nuclear and occasionally cytoplasmic Staining: antibody as precipitated red signal with a hematoxylin purple nuclear count

Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Analysis of FFPE prostate tissue using AKT1 phospho Ser473 antibody (left panel). Negative control (right panel). Antigen retrieval with heat and pressure in citrate buffer pH 6.2. AKT1 phospho Ser473 antibody at 20 ug/mL. Secondary antibody Streptavidin-HRP at 10 ug/mL. Hematoxylin nuclear counterstain (purple). Image using the Biotin format of this antibody.

Immunohistochemistry of Mouse anti-AKT pS473 antibody. Tissue: human prostate tissue. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 20 ug/mL for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear and occasionally cytoplasmic. Staining: AKT pS473 as precipitated red signal with hematoxylin purple nuclear counterstain.

Western Blot of Mouse Anti-AKTpS473 antibody. Lane 1: A431 cells. Lane 2: A431 cells stimulated for 15 min with EGF. Load: 35 ug per lane. Primary antibody: AKTpS473 antibody at 1:400 for overnight at 4C. Secondary antibody: DyLight(TM)649 Conjugated Anti-AKT1 [p Ser473] Antibody (17F6.B11) at 1:10,000 for 45 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting overnight at 4C. Predicted/Observed size: 56kDa. Other band(s): none.

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Pm

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Biotin	NBP2-21678
DyLight 488	NBP1-77701

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NB600-590

AKT1 [p Ser473] Antibody - BSA Free

(3 Publications)

0.1 mg

NB600-590

Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry of AKT11 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 1:100 for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.

Western Blot: AKT1 [p Ser473] Antibody [NB600-590] - Lane 1: nuclear extract from cells infected with adenovirus expressing nuclear-targeted AKT kinase. Load: 35 ug per lane. Primary antibody: AKT pS473 antibody at 1:200 dilution for overnight at 4C. Secondary antibody: IRDye800 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 56 kDa for AKT pS473. Other band(s): unspecific.

<b>Biological Strategies Validation. </b>AKT1-[p-Ser473]-Antibody-Western-Blot-NB600-590-img0012.jpg

Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry of AKT1 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 100 dilution for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.

Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry at higher magnification of AKT1 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 100 dilution for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.

Dot Blot: AKT1 [p Ser473] Antibody [NB600-590]

Dot Blot of Rabbit Anti-AKT pS473 Antibody. Dilutions in Columns: (1) 100ng, (2) 33.33ng, (3) 11.11ng, (4) 3.7ng, (5) 1.23ng. Tested BSA Peptide Reactivity in Rows: (A) AKT1-BSA, (B) AKT1 pT308-BSA, (C) AKT1 S473-BSA, (D) AKT1 pS473-BSA, (E) CDC27 T244-BSA, (F) CDC27 pT244-BSA, (G) BSA control. Primary Antibody: Anti-AKTpS473 at 1ug/mL overnight at 2-8C. Secondary Antibody: Goat anti-Rabbit IgG HRP

Immunohistochemistry of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Immunohistochemistry at higher magnification of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Western Blot of Rabbit anti-AKT pS473 antibody. Lane 1: AKT1 Recombinant Protein

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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AF1775

AKT1 Antibody [Unconjugated]

(5 Publications)

50 μg

AF1775

HAF008). A specific band for Akt1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, TS1 mouse helper T cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat Akt1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1775) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # <a class=

HAF008). A specific band for Akt1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, TS1 mouse helper T cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat Akt1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1775) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Akt1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

HAF008). A specific band for Akt1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows recombinant human Akt1, Akt2, and Akt3 (1 ng/lane). PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat Akt1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1775) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # <a class=

HAF008). A specific band for Akt1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Akt1 at approximately 63 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat Akt1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1775). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

,

Applications: ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF1775-SP

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NBP3-21578

AKT [p Ser473] Antibody (SR2061) - BSA Free

100 ul

NBP3-21578

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: AKT [p Ser 473] Antibody (SR2061) [NBP3-21578] - Immunofluorescent analysis of NIH/3T3 cells treated with PDGF, using NBP3-21578.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP3-21578-50ul

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NBP1-69924

AKT1 [p Thr308] Antibody (18F3.H11) - BSA Free

0.1 mg

NBP1-69924

Immunohistochemistry-Paraffin: AKT1 [p Thr308] Antibody (18F3.H11) [NBP1-69924] - IHC analysis. Detection of phosphorylated AKT phospho Thr308 in human FFPE brain cerebellum tissue (40X). AKT1 phospho Thr308 antibody was used at 20 ug/mL. The image shows strong staining of Purkinje neurons. Red: AKT1 pT308. Purple: Hematoxylin nuclear counterstain.

Western Blot: AKT1 [p Thr308] Antibody (18F3.H11) [NBP1-69924] - WB analysis using the Biotin conjugate of AKT1 pT308 antibody. Detection of Lane 1: GST tagged AKT1 active recombinant protein. Lane 2: GST tagged AKT1 inactive recombinant protein. Load: 25 ng per lane. AKT pT308 Biotin conjugated antibody at 1:1000 overnight at 4C. Secondary antibody: HRP Streptavidin.

Immunocytochemistry/Immunofluorescence: AKT1 [p Thr308] Antibody (18F3.H11) [NBP1-69924] - ICC/IF analysis using the DyLight 488 conjugate of AKT1 pT308, NBP1-69924. Histone detection using DyLight 488 conjugate (green). Anti-Tubulin was detected using DyLight 549 conjugate (red). Nuclei were counter-stained using DAPI (blue).

ELISA: AKT1 [p Thr308] Antibody (18F3.H11) [NBP1-69924] - ELISA of AKT1 phospho Thr308 Biotin conjugated antibody. Antigen: Unconjugated AKT1 phospho Thr308 and AKT1 non-phospho Thr308. Coating amount: 0.1 ug per well. Primary antibody: AKT1 phospho Thr308 Biotin conjugated antibody at 5 ug/mL. Dilution series: 3-fold. Mid-point concentration: 5 ng/mL AKT1 phospho Thr308 Biotin conjugated antibody. Secondary antibody: Peroxidase streptavidin secondary antibody at 1:10,000. Substrate: TMB

<b>Biological Strategies Validation. </b>Western Blot of Mouse anti-AKT pT308 antibody. Lane 1: non-phosphorylated AKT in untreated NIH/3T3 cells

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , , ,

Applications: , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Biotin	NBP2-21679
DyLight 488	NBP1-77702

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

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Clonality

Clone

Images Type

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Species Glossary

AKT1 Products By Gene Name

AKT [p Ser473] Antibody [Unconjugated] - Pan Specific

AKT Antibody (281046) [Unconjugated] - Pan Specific

AKT1 [p Ser473] Antibody (104A282) - BSA Free

AKT [p Ser473] Antibody (545007) [Unconjugated] - Pan Specific

AKT1 Antibody (OTI4D6)

AKT1 Antibody (3A3)

AKT1 [p Thr308] Antibody (658320) [Unconjugated]

AKT1 [p Ser473] Antibody (17F6.B11) - BSA Free

AKT1 [p Ser473] Antibody - BSA Free

AKT1 Antibody [Unconjugated]

AKT [p Ser473] Antibody (SR2061) - BSA Free

AKT1 [p Thr308] Antibody (18F3.H11) - BSA Free