AKT [p Ser473] Antibody (545007) - 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.

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

AKT [p Ser473] Antibody (545007) - 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: Protein A or G purified from hybridoma culture supernatant
• 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
• Purity: Protein A or G purified from hybridoma culture supernatant
• 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) - 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)(4)

Publications using MAB887

• 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;11(3):. 2022 [PMID: 35159188] (Western Blot, Human)
• 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;12(1):56. 2021 [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;21(4):442-454. 2020 [PMID: 32152508] (Flow Cytometry, Human)
• 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;34(2):197-203. 2017 [PMID: 28566814] (Infared Microscopy, Human)

Bioinformatics

• Gene Symbol: AKT1