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

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.

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 # 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.

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.

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.

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
• Applications: WB, Simple Western
• Clone: 545007
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

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

• Immunogen: Phosphopeptide containing the human Akt (S473) site
• Modification: p Ser473
• Specificity: Detects human and mouse Akt1, Akt2 and Akt3, when phosphorylated at S473, S474 and S472, respectively.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: AKT1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Simple Western 2 ug/mL
  • Western Blot 0.2 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

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

• RAC-ALPHA
• AKT serine/threonine kinase 1
• Akt
• PKB
• PKB-ALPHA
• PRKBA
• RAC

Background

Akt, also known as protein kinase B (PKB), is a central kinase in such diverse cellular processes as glucose uptake, cell cycle progression, and apoptosis. Three highly homologous members define the Akt family: Akt1 (PKB alpha ), Akt2 (PKB beta ), and Akt3 (PKB gamma ). All three Akts contain an amino-terminal pleckstrin homology domain, a central kinase domain, and a carboxyl-terminal regulatory domain. Akt1 is the most widely expressed family member and is frequently activated in a number of carcinomas, including breast, prostate, lung, pancreatic, liver, ovarian, and colorectal cancer. Akt1 is activated in a multistep process that involves the sequential phosphorylation of Thr450 by JNK kinases, Thr308 by PDK1, and Ser473 by PDK2 or mTORC2. Activated Akt1 phosphorylates a wide variety of cytosolic, nuclear, and mitochondrial substrates. Human Akt1 shares 98% aa sequence identity with mouse and rat Akt1.

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.

Publications for AKT Antibody (MAB887)(12)

Publications using MAB887

• Khorramdelazad, H;Bagherzadeh, K;Rahimi, A;Darehkordi, A;Najafi, A;Karimi, M;Khoshmirsafa, M;Hassanshahi, G;Safari, E;Falak, R; A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer Cancer cell international 2025-01-05 [PMID: 39757159] (Western Blot, Mouse)
• Klaudia Brodaczewska, Aleksandra Majewska, Aleksandra Filipiak-Duliban, Claudine Kieda Pten knockout affects drug resistance differently in melanoma and kidney cancer Pharmacol Rep 2023-09-06 [PMID: 37673853]
• Liang Z, Zhang Z, Zhang Q et al. The proprotein convertase furin regulates the development of thymic epithelial cells to ensure central immune tolerance iScience 2022-10-21 [PMID: 36274943]
• Shinji Sugita, Yoshifumi Naito, Li Zhou, Hongli He, Qi Hao, Atsuhiro Sakamoto et al. Hyaluronic acid restored protein permeability across injured human lung microvascular endothelial cells FASEB BioAdvances 2022-09-01 [PMID: 36089980]
• Majewska A, Brodaczewska K, Filipiak-Duliban A et al. miRNA Pattern in Hypoxic Microenvironment of Kidney Cancer-Role of PTEN Biomolecules 2022-05-11 [PMID: 35625614]
• K Kwa?niak, J Czarnik-Kw, K Malysheva, K Pogoda, O Korchynsky, P Rybojad, B Karczmarek, J Tabarkiewi LncCDH5-3:3 Regulates Apoptosis, Proliferation, and Aggressiveness in Human Lung Cancer Cells Cells, 2022-01-23;11(3):. 2022-01-23 [PMID: 35159188] (Western Blot, Human)
• Rodrigo Azevedo Loiola, Miguel García-Gabilondo, Alba Grayston, Paulina Bugno, Agnieszka Kowalska, Sophie Duban-Deweer et al. Secretome of endothelial progenitor cells from stroke patients promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage Stem Cell Research & Therapy 2021-10-26 [PMID: 34702368]
• Ö Boyac?o?lu, E Bilgiç, C Varan, E Bilensoy, E Nemutlu, D Sevim, Ç Kocaefe, P Korkusuz ACPA decreases non-small cell lung cancer line growth through Akt/PI3K and JNK pathways in vitro Cell Death & Disease, 2021-01-11;12(1):56. 2021-01-11 [PMID: 33431819] (Western Blot, Human)
• B Diskin, S Adam, MF Cassini, G Sanchez, M Liria, B Aykut, C Buttar, E Li, B Sundberg, RD Salas, R Chen, J Wang, M Kim, MS Farooq, S Nguy, C Fedele, KH Tang, T Chen, W Wang, M Hundeyin, JAK Rossi, E Kurz, MIU Haq, J Karlen, E Kruger, Z Sekendiz, D Wu, SAA Shadaloey, G Baptiste, G Werba, S Selvaraj, C Loomis, KK Wong, J Leinwand, G Miller PD-L1 engagement on T cells promotes self-tolerance and suppression of neighboring macrophages and effector T cells in cancer Nat. Immunol., 2020-03-09;21(4):442-454. 2020-03-09 [PMID: 32152508] (Flow Cytometry, Human, Mouse)
• Huiguo Shan, Xuefeng Zhou, Chuanjun Chen MicroRNA‑214 suppresses the viability, migration and invasion of human colorectal carcinoma cells via targeting transglutaminase 2 Molecular Medicine Reports 2019-06-03 [PMID: 31173203]
• MT Lane, TJ Herda, AC Fry, MA Cooper, MJ Andre, PM Gallagher Endocrine responses and acute mTOR pathway phosphorylation to resistance exercise with leucine and whey Biol Sport, 2017-01-20;34(2):197-203. 2017-01-20 [PMID: 28566814] (Infared Microscopy, Western Blot, Human)
• HUA WANG, JINLIN WU 17 beta -estradiol suppresses hyperoxia-induced apoptosis of oligodendrocytes through paired-immunoglobulin-like receptor B Molecular Medicine Reports 2016-03-01 [PMID: 26821234]

Bioinformatics

• Gene Symbol: AKT1