AKT1 [p Ser473] Antibody (17F6.B11)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-69923

Novus Biologicals, part of Bio-Techne
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NBP1-69923
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey

Cited:

Human, Mouse

Applications

Validated:

Dot Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 17F6.B11

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all Akt1 Antibodies »

Product Specifications

Immunogen

AKT1 [p Ser473] Antibody (17F6.B11) was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT11 protein, followed by hybridoma development. (Uniprot: P31749)

Reactivity Notes

A BLAST analysis was used to suggest cross-reactivity with AKT1 pS473 from human, mouse, rat and chimpanzee sources based on 100% homology with the immunizing sequence. Cross-reactivity with AKT1 from other sources has not been determined. Cross-reactivity with AKT2 and AKT3 has not been determined.

Modification

p Ser473

Specificity

This phospho specific monoclonal antibody is specific for phosphorylated human and mouse AKT protein at S473. A BLAST analysis was used to suggest cross-reactivity with AKT pS473 from human, mouse, rat and chimpanzee sources based on 100% homology with the immunizing sequence. Cross-reactivity with AKT from other sources has not been determined. Cross-reactivity with AKT2 and AKT3 has not been determined.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Description

This antibody Monoclonal Antibody was purified from concentrated tissue culture supernate by Protein A chromatography

Store this antibody at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for AKT1 [p Ser473] Antibody (17F6.B11)

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - A431 cells. Panel A: serum starved, unstimulated cells. Panel B: serum starved, EGF stimulated for 15 mins. A massive increase in AKT-pS473 activation, as measured by intensity signal, peaked at 15 minutes and was associated with depolymerized tubulin. Panel A shows STED data (AKT-pS473, red channel) collected simultaneously with confocal signal (alpha-tubulin, green channel). Upon stimulation of cells with EGF, a rapid activation of AKT is observed (Panel B) along with a coincident change in the tubulin organization (yellow signal), as well as an extensive cell shape-change (cell membrane folding) and accumulation of AKT pS473 at the cell periphery.
Flow Cytometry: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Analysis using the DyLight 488 conjugate of AKT1 phospho Ser473 antibody. Image shows anti-histone detection using a DyLight 488 conjugate (green). Anti-Tubulin was detected using a DyLight 549 conjugate (red). Nuclei were counter-stained using DAPI (blue).
Western Blot: AKT1 [p Ser473] Antibody (17F6.B11) [NBP1-69923] - Western Blot of AKT1 [p Ser473] antibody (17F6.B11). A: Lane 1) PDGF stimulated NIH 3T3 cells [10ul]. Lane 2) NIH 3T3 cells [10ul]. Lane 3) Hela whole cell lysate [10ul] (weak signal). B: Lane 4) GST negative control protein [100ng]. Lane 5) GST negative control protein [25ng]. Lane 6) AKT 1 recombinant protein [100ng]. Lane 7) AKT 1 recombinant protein [25ng].Block: 5% BSA overnight at 4C.Primary antibody: Monoclonal anti-AKT antibody used at 1:1000 for overnight at 4C.Secondary antibody: HRP Conjugated goat anti-mouse 1:25K for 45 min at RT.Detection: TMB for 20 minutes, rinsed with deionized water, dried and scanned on conventional flatbed scanner.
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Applications for AKT1 [p Ser473] Antibody (17F6.B11)

Application
Recommended Usage

ELISA

1:20000

Flow Cytometry

1:10-1:1000

Immunocytochemistry/ Immunofluorescence

1:500-1:3000

Immunohistochemistry

20 ug/ml

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:500-1:3000
Application Notes
This product is tested in ELISA, immunofluorescence, immunohistochemistry, and western blotting. Expect a band approximately 56 kDa in size corresponding to phosphorylated AKT protein by western blotting in the appropriate cell lysate or extract. This phospho-specific monoclonal antibody reacts with human and mouse AKT pS473 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. Specific conditions for reactivity should be optimized by the end user. For immunohistochemistry use formalin-fixed paraffin-embedded sections. No pre-treatment of sample is required.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at -20C. 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

Gene Symbol

AKT1

UniProt

P31749

Additional Akt1 Products

Product Documents for AKT1 [p Ser473] Antibody (17F6.B11)

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Product Specific Notices for AKT1 [p Ser473] Antibody (17F6.B11)

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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