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

Concentration

1.0 mg/ml

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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.

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.
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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

Product Datasheets

Certificate of Analysis

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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.

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