Key Product Details

Validated by

Independent Antibodies, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Rabbit

Cited:

Human, Mouse, Rat, Rabbit

Predicted:

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

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 104A282

Format

BSA Free

Concentration

1 mg/ml

View all Akt1 Antibodies »

Product Specifications

Immunogen

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

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 31092832). Rabbit reactivity reported in scientific literature (PMID: 32936958)

Modification

p Ser473

Specificity

Clone 104A282 detects specifically the Ser473 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) - BSA Free

Western Blot: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - 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.
Western Blot: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - WB of phospho AKT using phospho AKT 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. Image using the Azide and BSA Free form of this antibody.
Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - 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.
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Applications for AKT1 [p Ser473] Antibody (104A282) - BSA Free

Application
Recommended Usage

Immunohistochemistry

1:200 - 1:250

Immunohistochemistry-Frozen

reported by customer review

Immunohistochemistry-Paraffin

1:200 - 1:250
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using NB100-56749 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 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

UniProt

P31749

Additional Akt1 Products

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

Product Datasheets

Certificate of Analysis

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Product Specific Notices for AKT1 [p Ser473] Antibody (104A282) - 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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