Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Rabbit

Predicted:

Amphibian (100%), Chicken (100%), Zebrafish (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 104A282

Format

Azide and BSA Free
View all Akt1 Primary Antibodies »

Product Specifications

Immunogen

This AKT1 phospho Ser473 monoclonal antibody was raised against a synthetic peptide containing phosphorylated serines at amino acid residues 473 of human Akt1.

Modification

p Ser473

Specificity

Clone 104A282 detects specifically the Ser 473 phosphorylated form of AKT1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

55.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

Novus Biologicals Mouse AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free (NB100-56749) is a monoclonal antibody validated for use in IHC and WB. Anti-AKT1 Antibody: Cited in 6 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

Western Blot: AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free [NBP2-27399] - Total protein from mouse 3T3 cells treated with and without PDGF (50 ng/mL) for the indicated times was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-AKT1 (NBP2-01725) and 2 ug/mL pS473 AKT1 in 1% BSA in TBST and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Note the detection of phosphorylated AKT1 in response to PDGF treatment compared to total AKT1 protein. Image using the standard format of this product.
Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free [NBP2-27399] - IHC analysis of an FFPE human breast carcinoma tissue section @ 1:250 of pSer473 AKT1 antibody (clone 104A282) on a Bond Rx autostainer (Leica Biosystems). Assay: 20 mins of heat induced antigen retrieval (HIER) with 10 mM sodium citrate buffer (pH 6.0) & endogenous peroxidase quenching using peroxide block. Sections incubated with primary antibody for 30 mins. Bond Polymer Refine Detection (Leica Biosystems) & DAB used for signal detection following counterstaining with hematoxylin. Whole slide scanning & capturing of representative images (20X) performed using Aperio AT2 (Leica Biosystems). Antibody generated diffused cytoplasmic staining of phospho-AKT (Ser-473) in the cancer cells as well as the stromal cells. Some cancer cells depicted nuclear staining also. Staining was performed by Histowiz. Image using the standard format of this product.
Western Blot: AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free [NBP2-27399] - WB analysis of phospho AKT using AKT1 phospho Ser473 antibody at 2 ug/mL against untreated (lane 1) and PDGF treated (lane 2) NIH-3T3 lysate. HRP-conjugated secondary antibody and ECL substrate solution were used for this test.
Immunohistochemistry: AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free [NBP2-27399] - 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. Image using the standard format of this product.

Applications for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

Application
Recommended Usage

Western Blot

1 - 2 ug/mL

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Akt1

AKT (also known as protein kinase B (PKB) and RAC (related to A and C kinases)) is a critical intracellular serine/threonine kinase that translates signals from extracellular stimuli including growth factors, cytokines and neurotransmitters (1). AKT signaling plays critical roles in cell growth, proliferation, survival and differentiation (1). It is also involved in organogenesis, angiogenesis and metabolism. Three mammalian AKT isoforms have been identified. The AKT pathway can be activated by any of the three members who share a high level of protein homology but are independently encoded by AKT1 (PKB alpha; 14q32.32), AKT2 (PKB beta; 19q13.2), or AKT3 (PKB gamma; 1q44) (1, 2). Each AKT family member contains an N-terminal pleckstrin homology (PH) domain, a central kinase domain, and a C-terminal regulatory domain. AKT mediates many of the downstream events of phosphatidylinositol 3-kinase (PI3-K), a lipid kinase activated by growth factors, cytokines and insulin. PI3-K recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. AKT has two main phosphorylation sites (Ser473 and Thr308, predicted molecular weight 56 kDa) (3, 4). Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus including mammalian target of rapamycin (mTOR).

The main function of AKT is to control inhibition of apoptosis and promote cell proliferation. Survival factors can activate AKT Ser473 and Thr308 phosphorylation sites in a transcription-independent manner, resulting in the inactivation of apoptotic signaling transduction through the tumor suppressor PTEN, an antagonist to PI3-K (5). PTEN exerts enzymatic activity as a phosphatidylinositol-3,4,5-trisphosphate (PIP3) phosphatase, opposing PI3K activity by decreasing availability of PIP3 to proliferating cells, leading to overexpression and inappropriate activation of AKT noted in many types of cancer.

AKT1 function has been linked to overall physiological growth and function (2). AKT1 has been correlated with proteus syndrome, a rare disorder characterized by overgrowth of various tissues caused by a mosaic variant in the AKT1 gene in humans.

AKT2 is strongly correlated with Type II diabetes, including phenotypes of insulin resistance, hyperglycemia and atherosclerosis (2, 6).

The function of AKT3 is specifically associated to brain development, where disruptions to AKT3 are correlated with microcephaly, hemimegalencephaly, megalencephaly and intellectual disabilities (2).

References

1. Ersahin, T., Tuncbag, N., & Cetin-Atalay, R. (2015). The PI3K/AKT/mTOR interactive pathway. Mol Biosyst, 11(7), 1946-1954. doi:10.1039/c5mb00101c

2. Cohen, M. M., Jr. (2013). The AKT genes and their roles in various disorders. Am J Med Genet A, 161a(12), 2931-2937. doi:10.1002/ajmg.a.36101

3. Georgescu, M. M. (2010). PTEN Tumor Suppressor Network in PI3K-Akt Pathway Control. Genes Cancer, 1(12), 1170-1177. doi:10.1177/1947601911407325

4. Mishra, P., Paital, B., Jena, S., Swain, S. S., Kumar, S., Yadav, M. K.,... Samanta, L. (2019). Possible activation of NRF2 by Vitamin E/Curcumin against altered thyroid hormone induced oxidative stress via NFkB/AKT/mTOR/KEAP1 signalling in rat heart. Sci Rep, 9(1), 7408. doi:10.1038/s41598-019-43320-5

5. Wedel, S., Hudak, L., Seibel, J. M., Juengel, E., Oppermann, E., Haferkamp, A., & Blaheta, R. A. (2011). Critical analysis of simultaneous blockage of histone deacetylase and multiple receptor tyrosine kinase in the treatment of prostate cancer. Prostate, 71(7), 722-735. doi:10.1002/pros.21288

6. Rotllan, N., Chamorro-Jorganes, A., Araldi, E., Wanschel, A. C., Aryal, B., Aranda, J. F.,... Fernandez-Hernando, C. (2015). Hematopoietic Akt2 deficiency attenuates the progression of atherosclerosis. Faseb j, 29(2), 597-610. doi:10.1096/fj.14-262097

Long Name

v-Akt Murine Thymoma Viral Oncogene Homolog 1

Alternate Names

PKB alpha, PRKBA, RAC-alpha

Entrez Gene IDs

207 (Human); 11651 (Mouse)

Gene Symbol

AKT1

Additional Akt1 Products

Product Documents for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

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.

Citations for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for AKT1 [p Ser473] Antibody (104A282) - Azide and BSA Free

Showing  1 - 5 of 5 FAQs Showing All

  • Q: Do your HRP-conjugated antibodies contain sodium azide?

    A: No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: 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).

  • Q: 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.

    A: At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.

  • Q: What is the molecular weight of your antibodies?

    A: All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

  • Q: Why are many of your antibodies formulated with sodium azide and BSA?

    A: Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.

  • Q: Do your HRP-conjugated antibodies contain sodium azide?

    A: No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: 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).

  • Q: 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.

    A: At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.

  • Q: What is the molecular weight of your antibodies?

    A: All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

  • Q: Why are many of your antibodies formulated with sodium azide and BSA?

    A: Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.

  • Q: Do your HRP-conjugated antibodies contain sodium azide?

    A: No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: 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).

  • Q: 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.

    A: At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.

  • Q: What is the molecular weight of your antibodies?

    A: All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

  • Q: Why are many of your antibodies formulated with sodium azide and BSA?

    A: Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.

  • Q: Do your HRP-conjugated antibodies contain sodium azide?

    A: No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: 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).

  • Q: 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.

    A: At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.

  • Q: What is the molecular weight of your antibodies?

    A: All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

  • Q: Why are many of your antibodies formulated with sodium azide and BSA?

    A: Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.

  • Q: Do your HRP-conjugated antibodies contain sodium azide?

    A: No. None of our HRP-conjugated antibodies contain sodium azide as this agent inhibits the activity of HRP.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: 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).

  • Q: 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.

    A: At the moment we do not have an AKT1 antibody that definitively does not react with either AKT2 or AKT3.

  • Q: What is the molecular weight of your antibodies?

    A: All IgG antibodies are approximately 150 kDa (each heavy chain is about 50 kDa and each light chain is about 25 kDa).

  • Q: Why are many of your antibodies formulated with sodium azide and BSA?

    A: Sodium azide is a preservative which is added to prevent bacterial growth. BSA is added as a protein stabilizer.

Showing  1 - 5 of 5 FAQs Showing All
View all FAQs for Antibodies