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APAF-1 Antibody

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Western Blot: APAF1 Antibody [NB100-56074] - Analysis of Apaf 1 using this antibody. Jurkat: human T-cell leukemia cell line lysate. RS11-846: human B-cell leukemia cell line lysate. Human thymus: tissue lysate.
Western Blot: APAF1 Antibody [NB100-56074] - Analysis of Apaf 1 using this antibody. Jurkat: human T-cell leukemia cell line lysate. RS11-846: human B-cell leukemia cell line lysate. Human thymus: tissue lysate.
Immunocytochemistry/ Immunofluorescence: APAF1 Antibody [NB100-56074] - Formalin-fixed, paraffin-embedded sections from a human brain tumor tissue microarray stained for Apaf 1 expression using NB100-56074 at 1:2000. A ...read more
Immunohistochemistry: APAF-1 Antibody [NB100-56074] - APAF1 Antibody [NB100-56074] - Formalin-fixed, paraffin-embedded tissue sections of human brain tumors stained for Apaf 1 expression using NB100-56074 at 1:2000. A. ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, MaSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-Fr, IHC-P, IP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated

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APAF-1 Antibody Summary

Immunogen
Full-length recombinant human Apaf1 protein was used as immunogen.
Specificity
Apaf 1
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
APAF1
Purity
Unpurified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
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Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:2000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:1000-1:5000
  • Immunoprecipitation 1:50-1:200
Control
APAF-1 Knockout HeLa Cell Lysate
Publications
Read Publication using
NB100-56074 in the following applications:

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
Neat whole antisera
Preservative
0.05% Sodium Azide
Purity
Unpurified

Alternate Names for APAF-1 Antibody

  • APAF1
  • APAF-1
  • APAF-1apoptotic protease activating factor 1
  • apoptotic peptidase activating factor 1
  • apoptotic peptidase activating factor
  • apoptotic protease activating factor
  • apoptotic protease-activating factor 1
  • CED4
  • DKFZp781B1145
  • KIAA0413

Background

Apaf 1 (apoptosis protease-activating factor-1) is a key regulator of the mitochondrial apoptotic pathway (reviewed in Campioni et al, 2005), being the central element of the multimeric apoptosome. The apoptosome consists of cytochrome c, procaspase-9 and seven Apaf 1 monomers, and is considered to be core apoptotic machinery that executes mitochondria-dependent apoptosis. Cytochrome c, normally compartmentalized in the mitochorndria, is released into the cytoplasm following apoptotic stimuli. Apaf 1 binds cytochrome c in the cytoplasm and in the presence of dATP/ATP forms the apoptosome. The apoptosome binds procaspase-9 and promotes its autocatalytic activation. Active caspase-9, in turn, activates downstream caspases including 3, 6, and 7 contributing to the proteolytic caspase activation cascade which leads to cell death. Apaf 1 is an approx. 130 kDa protein. Alternative Apaf 1 splicing isoforms of Apaf 1 have been identified in mammalian cells, each having either or both an 11 amino acid insertion in the N-terminus and an additional WD40 repeat (43 aa) in the C-terminus; please refer to AceView of NCBI (http://www.ncbi.nih.gov/IEB/Research/Acembly/) for detailed information. Additionally, Apaf 1 can be proteolytically processed by active caspase-3 during apoptosis resulting in an Apaf 1 cleavage fragment of ~84 kDa (Lauber et al. 2001).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Immunohistochemistry: Caspase-9 Antibody [NB100-56118] - Mice with two different spontaneous mutant alleles of Sharpin show variations in onset of CPDM phenotype. However, the onset is much more rapid in Sharpincpdm-Dem mice as evidenced by antibody detection of cleaved CASPASE 9 (T) when compared to Sharpincpdm mutants (R) or WT mice (Q,S).. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0085666) licensed under a CC-BY licence.
Immunohistochemistry: Caspase-9 Antibody [NB100-56118] - Autophagy-deficient mice showed increased activity of apoptosis and necroptosis. Increased the expression of Caspase-9 in 12-week-old Atg7deltapan mice. Caspase-9 quantitation and representative IF microphotographs of Atg7F/F (n=5) and Atg7deltapan (n=5) pancreatic tissue stained for DAPI (blue) and Caspase-9 (green) (1/1000, scale bar=50?um. Image collected and cropped by CiteAb from the following publication (http://www.nature.com/doifinder/10.1038/cddis.2017.313), licensed under a CC-BY licence.
Immunohistochemistry-Paraffin: Caspase-9 Antibody [NB100-56118] - Analysis of Capase-9 in mouse transplanted lung tumor section using anti-Caspase-9 antibody. Image from verified customer review.
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Western Blot: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - Analysis using the biotin conjugate of NB100-56503. Detection of Cytochrome C in 15 mg of HeLa cell lysate using NB100-55775 at 1:1000. A 15 kDa band is detected.
Immunocytochemistry/Immunofluorescence: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - p73 was detected in immersion fixed Hela human cell line using NB100-56503 at 25 ug/ml for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Staining was observed in the cytoplasm and mitochondria.
Immunohistochemistry-Paraffin: Cytochrome c Antibody (7H8.2C12) [NB100-56503] - Cytochrome C was detected in immersion fixed paraffin-embedded sections of human heart using anti-human mouse monoclonal antibody (Catalog # NB100-56503) at 1:200 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue).Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
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Caspase-3 Antibody
(31A1067) - (Pro and ...
NB100-56708
Western Blot: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Image of Caspase-3 Antibody (31A1067) - (Pro and Active). Whole cell protein from Jurkat cells treated with and without 2 uM staurosporine as indicated was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 5 ug/ml anti-Caspase 3 in 1% milk, and detected with an anti-mouse HRP secondary antibody using a Femto sensitivity chemiluminescence reagent. Note the detection of both pro-caspase 3 at 35 kDa and the cleaved active caspase 3 at 15-17 kDa.
Immunohistochemistry-Paraffin: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Tissue section of human spleen using 1:200 dilution of Caspase-3 antibody (clone 31A1067). The staining was developed with HRP labeled anti-mouse IgG secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Caspase 3 antibody generated primarily a specific cytoplasmic staining in a subset of spleenocytes with some nuclear signal in a few cells.
Simple Western: Caspase-3 Antibody (31A1067) - (Pro and Active) [NB100-56708] - Simple Western lane view shows a specific band for Caspase-3 Antibody (31A1067) - (Pro and Active) in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.
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NB100-56098
Western Blot: Bcl-2 Antibody [NB100-56098] - Analysis of Bcl-2 in whole cell lysate from Daoy cells. Cells were transfected with (1) scrambled control siRNA or (2) Bcl-2 siRNA. Image from verified customer review.
Western Blot: Bcl-2 Antibody [NB100-56098] - Analysis of whole cell lysate (WCL) of Jurkat cells using anti-Bcl-2 antibody (NB100-56098) at 1:1000 dilution. HRP conjugated goat anti-rabbit IgG (H+L) cross adsorbed secondary antibody was used with ECL substrate for the detection of Bcl2 antibody bound to the blotted protein. This Bcl2 antibody detected a specific band at expected molecular weight marker position of Bcl2 protein (26kDa).
Immunohistochemistry-Paraffin: Bcl-2 Antibody [NB100-56098] - Analysis of a FFPE human breast carcinoma tissue section using 1:1000 dilution of Bcl-2 antibody (NB100-56098) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM 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 BCL2 in the cancer cells with relatively intense signal in their nuclear membranes. Staining was performed by Histowiz.
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Western Blot: Bcl-xL Antibody [NB100-56104] - WB analysis of tissue lysates from breast surgical tumor tissue biopsies (Lanes 1-7). Bcl-XS is not detected and variable amounts of Bcl-XL (~30 kDa) is detected. Lane 8. Recombinant Bcl-XL (positive control).
Western Blot: Bcl-xL Antibody [NB100-56104] - Analysis of Bcl-xL in Daoy whole cell lysate using anti-Bcl-xL antibody. Image from verified customer review.
Immunohistochemistry-Paraffin: Bcl-xL Antibody [NB100-56104] - Formalin-fixed paraffin-embedded sections of human epidermis stained for Bcl-xL expression. The bottom image was negative for Bcl-xL expression. The adjacent stratum spinosum and middle stratum granulosum layers were both very positive for Bcl-xL expression. The top stratum corneum consisting mainly of dead or peeling cells were both negative for Bcl-xL expression.
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Western Blot: Bax Antibody (6A7) [NBP1-28566] - rOPN administration elevated the expression of autophagy-related proteins while suppressing apoptosis in rat brain at 24 h after SAH. The effects of rOPN on expression levels of Bax, mean +/- SD is 1.006 +/- 0.321 in Sham group, 37.47 +/- 10.86 in SAH + Vehicle group, 23.83 +/- 8.143 in SAH + rOPN group, F = 33.13, in the left hemisphere of rat brain at 24 h after SAH. Sample size is 18, n = 6 per group. Data were presented as mean +/- SD. *P <.05, ***P <.001 vs Sham group; #P <.05, ##P <.01 vs SAH + Vehicle group Image collected and cropped by CiteAb from the following publication (https://onlinelibrary.wiley.com/doi/abs/10.1111/cns.13199) licensed under a CC-BY licence.
Immunohistochemistry-Frozen: Bax Antibody (6A7) [NBP1-28566] - Mouse retina with no pretreatment. IHC-F image submitted by a verified customer review
Western Blot: Bax Antibody (6A7) [NBP1-28566] - Analysis of lysates from human metastatic pancreatic cancer cell line L3.6pl using Bax Antibody (clone 6A7; Lot # B168-P780E). This image was submitted via reviews by a varified end user.
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Immunohistochemistry-Paraffin: p53 Antibody (PAb 240) [NB200-103] - p53 was detected in immersion fixed paraffin-embedded sections of human prostate cancer using anti-human mouse monoclonal antibody (Catalog # NB200-103) at 1:200 dilution overnight at 4C. Tissue was stained using the VisuCyte anti-mouse HRP polymer detection reagent (Catalog # VC001) with DAB chromogen (brown) and counterstained with hematoxylin (blue). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.
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Western Blot: XIAP Antibody - Unpurified [NB100-56183] - Analysis of XIAP in various tumor cell lines using this antibody at 1:2000.
Immunohistochemistry-Paraffin: XIAP Antibody - Unpurified [NB100-56183] - Human breast carcinoma using this antibody at 1:2000. A-A3, successively higher magnifications of the breast carcinoma tissue section. Hematoxylin-eosin counterstain.
Western Blot: XIAP Antibody - Unpurified [NB100-56183] - XIAP Antibody [NB100-56183] - Analysis of recombinant full-length IAP proteins using this antibody to XIAP at 1:2000. The data shows that the antibody recognizes only XIAP and not the other IAP proteins.
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Western Blot: FLASH Antibody [NB500-218] - Analysis of FLASH in NIH3T3 cells using this antibody at 1:500 dilution.
Species: Hu, Mu
Applications: WB
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Western Blot: Smac Antibody (78-1-118) [NB500-213] - HeLa whole cell extract lysates.
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TNF-alpha
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1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
Recombinant Human TNF-alpha (Catalog # 210‑TA) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25‑100 pg/mL.
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Fas/TNFRSF6/CD95
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Western blot shows Jurkat human acute T cell leukemia cell line treated with apoptosis inducer anti-Fas for the indicated times. PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BID at approximately 20 kDa (as indicated). For additional reference short (5 seconds, left panel) and long (60 seconds, right panel) exposures are shown. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for BID at approximately 29 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.
Western blot shows lysates of SVEC4‑10 mouse vascular endothelial cell line and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for BID at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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AIF Antibody
[Unconjugated]
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Western blot shows lysates of MCF‑7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, CTLL‑2 mouse cytotoxic T cell line, DA3 mouse myeloma cell line, and PC‑12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for AIF at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for AIF at approximately 69 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western blot shows lysates of HEK293T human embryonic kidney parental cell line and AIF knockout HEK293T cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1457) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for AIF at approximately 65 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockoutHEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
(6 Publications)
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC, KO
PARP Antibody
(194C1439) - Cleaved
NB100-56599
Western Blot: PARP Antibody (194C1439) - Cleaved [NB100-56599] - Analysis of cleaved PARP in staurosporine-treated Jurkat cells at various time points, antibody at 2 ug/mL. The band corresponding to cleaved PARP is only seen in the treated samples. Anti-mouse Ig HRP conjugate was used in this test.
Immunocytochemistry/Immunofluorescence: PARP Antibody (194C1439) - Cleaved [NB100-56599] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-PARP Antibody (194C1439) at 5 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: PARP Antibody (194C1439) - Cleaved [NB100-56599] - Analysis using 293T lysates expressing PARP. Image from verified customer review.
(8 Publications)
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Applications: WB, Flow, ICC/IF, Flow-IC
p21/CIP1/CDKN1A
Antibody (WA-1)
NBP2-29463
Immunohistochemistry-Paraffin: p21/CIP1/CDKN1A Antibody (WA-1 (HJ21)) [NBP2-29463] - Formalin-fixed, paraffin-embedded human bladder carcinoma stained with p21 Monoclonal Antibody (WA-1).
(14 Publications)
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APAF-1 Antibody
NB100-56074
APAF-1 Antibody
Species: Hu, Mu, Rt, Ma
Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P, IP
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Publications for APAF-1 Antibody (NB100-56074)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: IHC-P.


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Control Lysate(s)

APAF-1 Knockout HeLa Cell Lysate

Secondary Antibodies

 

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Caspase 9 - an important apoptosis marker
Caspases are essential mediators of programmed cell death and are needed for both the induction of apoptosis as well as for aiding the degradation of cellular structures. Initiator caspases (such as Caspase-9) sense and respond to various signals i...  Read full blog post.
Heat Shock Proteins: An Overview
Heat Shock Proteins (HSPs) are a ubiquitous group of molecular chaperone proteins that have evolved unique mechanisms, within their host cells, to facilitate survival in hostile environments such as heat, oxidative (hypoxia), pH and cold. Under permis...  Read full blog post.
The Role of the Caspase 3 Antibody in Apoptosis Research
The caspases are a group of cysteine protease enzymes essential to apoptosis, inflammation and necrosis. Caspase 3 has been identified as one of the key mediators of apoptosis.Human caspases form three distinct groups: cytokine activators, which are...  Read full blog post.
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Bioinformatics

Gene Symbol APAF1
Entrez
Uniprot