AKT1 Antibody (710022) - (E17K Mutation)

Western blot shows lysates of 293-EBNA human EBV-expressing embryonic kidney cell line either transfected with Akt1 (wild-type) or transfected with Akt1 (E17K Mutation). PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Akt1 (E17K Mutation) Monoclonal Antibody (Catalog # MAB6815) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Akt1 (E17K Mutation) at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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 Akt1 (panels A and B) or E17K mutated Akt1 (panels C and D) using Mouse Anti-Human Akt1 (E17K Mutation) Monoclonal Antibody (Catalog # MAB6815) and Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055). 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.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clone: 710022
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

AKT1 Antibody (710022) - (E17K Mutation) Summary

• Immunogen: Synthetic peptide containing human Akt1 Ala5-Lys30 with Glu17Lys mutation
  Accession # P31749
• Specificity: Detects human AKT1 with Glu17Lys mutation in Western blots. In Western blots, no cross‑reactivity with 293‑EBNA human EBV-expressing embryonic kidney cell line transfectant expressing wild type AKT1 is detected.
• 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:
  • Immunocytochemistry 8-25 ug/mL
  • Western Blot 0.5 ug/mL

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 AKT1 Antibody (710022) - (E17K Mutation)

• AKT serine/threonine kinase 1
• AKT
• Akt1
• AKT1m
• CWS6
• EC 2.7.11
• EC 2.7.11.1
• PKB alpha
• PKB
• PKBMGC99656
• PRKBA
• Protein Kinase B Alpha
• Protein kinase B
• Proto-oncogene c-Akt
• rac protein kinase alpha
• RAC
• RAC-alpha serine/threonine-protein kinase
• RAC-alpha
• RAC-PK-alpha
• RACPKB-ALPHA
• Serine-Threonine Protein Kinase
• v-akt murine thymoma viral oncogene homolog 1
• V-Akt Murine Thymoma Viral Oncogene-Like Protein 1

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. Substitution of glutamic acid with lysine at amino acid 17 (E17K) occurs in multiple malignancies and results in constitutive Akt1 activation and cell transformation. Within aa 5‑30, human Akt1 shares 100% aa sequence identity with mouse and rat Akt1.

1. Carpten, J.D. et al. (2007) Nature 448:439.
2. Bleeker, F.E. et al. (2008) Oncogene 27:5648.
3. Brugge, J. et al. Cancer Cell 12:104.

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.

FAQs for AKT1 Antibody (MAB6815). (Showing 1 - 5 of 5 FAQ).

1. How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?
   • Use isotype-specific secondary antibodies if the primary antibodies are of different isotypes. You can also make direct conjugates of the primary antibodies by use of antibody labeling kits, dyes, or custom conjugations (please contact Technical Support for custom orders).
2. Why are many of your antibodies formulated with sodium azide and BSA?
   • Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.
3. Do your HRP-conjugated antibodies contain sodium azide?
   • No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.
4. I am looking for a antibody that recognizes human Akt1 but NOT Akt2 or 3, for Western blot analyses. I also want that antibody to recognize Akt1 regardless of its phosphorylated form.
   • At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.
5. What is the molecular weight of your antibodies?
   • All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

Bioinformatics

• Gene Symbol: AKT1
• Uniprot:
  • P31749()