AKT [p Ser473] Antibody (PSH04-44)

Novus Biologicals | Catalog # NBP3-32747

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH04-44 expressed in HEK293
View all Akt Primary Antibodies »

Product Specifications

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser473 of human AKT1. (Uniprot: P31749)

Modification

p Ser 473

Localization

Cytoplasm, Nucleus, Cell membrane.

Specificity

Detects endogenous levels of AKT1 only when phosphorylated at Ser473. This antibody also recognizes AKT2 and AKT3 when phosphorylated at the corresponding residues.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

56 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 AKT [p Ser473] Antibody (PSH04-44)

AKT [p Ser473] Antibody (PSH04-44)

Western Blot: AKT [p Ser473] Antibody (PSH04-44) [NBP3-32747]

Western Blot: AKT [p Ser473] Antibody (PSH04-44) [NBP3-32747] - Western blot analysis of AKT on different lysates with Rabbit anti-AKT antibody (NBP3-32747) at 1/2,000 dilution.

Lane 1: HEK-293 cell lysate
Lane 2: HEK-293 treated with 50uM LY294002 for 6 hours cell lysate

Lysates/proteins at 20 ug/Lane.

Predicted band size: 56 kDa
Observed band size: 56 kDa

Exposure time: 20 seconds; ECL;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32747) at 1/2,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
AKT [p Ser473] Antibody (PSH04-44) Western Blot: AKT [p Ser473] Antibody (PSH04-44) [NBP3-32747]

Western Blot: AKT [p Ser473] Antibody (PSH04-44) [NBP3-32747]

Western blot analysis on different lysates with NBP3-32747 at 1/2,000 dilution and pan AKT antibody at 1/2,000 dilution. Lane 1: MCF7 cell lysate Lane 2: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate Lane 3: SH-SY5Y cell lysate Lane 4: SH-SY5Y treated with 100ng/mL PDGF for 5 minutes cell lysate Lane 5: HEK-293 cell lysate Lane 6: HEK-293 treated with 50μM LY294002 for 6 hours cell lysate Lane 7: NIH/3T3 cell lysate Lane 8: NIH/3T3 treated with 100ng/mL PDGF for 5 minutes cell lysate Lane 9: C6 cell lysate Lane 10: C6 treated with 100ng/mL PDGF for 5 minutes cell lysate Lane 11: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, then the membrane treated with lambda pp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 56 kDa Observed band size: 56 kDa Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA for 1 hour at room temperature. NBP3-32747 at 1/2,000 dilution and pan AKT antibody (HA721870) at 1/2,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG [HRP] Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Applications for AKT [p Ser473] Antibody (PSH04-44)

Application
Recommended Usage

Western Blot

1:2000-1:5000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

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: Akt

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 neurotransmitters1. AKT signaling plays critical roles in cell growth, proliferation, survival and differentiation1. 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 56kD)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-K5. PTEN exerts enzymatic activity as a phosphatidylinositol-3,4,5-trisphosphate (PIP3) phosphatase, opposing PI3K activity by increasing 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 function2. 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 atherosclerosis2,6.

The function of AKT3 is specifically associated to brain development, where disruptions to AKT3 are correlated with microcephaly, hemimegalencephaly, megalencephaly and intellectual disabilities2.

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

Alternate Names

PKB, RAC

Gene Symbol

AKT1

Additional Akt Products

Product Documents for AKT [p Ser473] Antibody (PSH04-44)

Certificate of Analysis

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Product Specific Notices for AKT [p Ser473] Antibody (PSH04-44)

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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