AKT1 [p Ser473] Antibody - BSA Free

Novus Biologicals | Catalog # NB600-590

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Dot Blot

Cited:

Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Akt1 Primary Antibodies »

Product Specifications

Immunogen

AKT1 [p Ser473] Antibody was prepared by repeated immunizations in rabbits with a synthetic peptide corresponding to a C-terminus region near phospho Serine 473 of the human, mouse, rat and chicken AKT1 proteins conjugated to KLH. (Uniprot: P31749)

Reactivity Notes

This antibody is specific for phosphorylated human AKT1 pS473. Minimal reactivity occurs against non-phosphorylated AKT. Reactivity against AKT1 from other species may occur but has not yet been tested.

Modification

p Ser473

Localization

Cytoplasmic and Nuclear

Specificity

This antibody is specific for phosphorylated human AKTpS473. Minimal reactivity occurs against non-phosphorylated AKT. Reactivity against AKT from other species may occur but has not yet been tested.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This product was prepared from monospecific antiserum by immunoaffinity chromatography using a phospho peptide coupled to agarose beads followed by solid phase adsorption(s) against a non-phospho peptide and non-specific peptide to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum

Store vial 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 - BSA Free

Western Blot: AKT1 [p Ser473] Antibody [NB600-590] - Western Blot of AKT11 [p Ser473] antibody. Lane 1: AKT11 Recombinant Protein. Lane 2: AKT11 Mutant Human Recombinant Protein.Lane 3: AKT11 (phosphatase treated) Human Recombinant Protein. Load: 50 ng per lane.Primary antibody: AKT1 pS473 antibody at 1:1,000 for overnight at 4C.Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT.Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT.Predicted/Observed size: ~56 kDa for AKT1pS473.
Immunocytochemistry/ Immunofluorescence: AKT1 [p Ser473] Antibody [NB600-590]

Immunocytochemistry/ Immunofluorescence: AKT1 [p Ser473] Antibody [NB600-590]

Immunocytochemistry/Immunofluorescence: AKT1 [p Ser473] Antibody [NB600-590] - Cardiomyocytes infected with adenovirus expressing wild-type AKT. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 1:40 for 1 h at RT. Secondary antibody: texas-red conjugated rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: AKT pS473 is nuclear. Staining: AKT pS473 as green fluorescent signal with texas-red conjugated phalloidin (red) to label filamentous actin.
Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry of AKT11 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 1:100 for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.
Western Blot: AKT1 [p Ser473] Antibody [NB600-590]

Western Blot: AKT1 [p Ser473] Antibody [NB600-590]

Western Blot: AKT1 [p Ser473] Antibody [NB600-590] - Lane 1: nuclear extract from cells infected with adenovirus expressing nuclear-targeted AKT kinase. Load: 35 ug per lane. Primary antibody: AKT pS473 antibody at 1:200 dilution for overnight at 4C. Secondary antibody: IRDye800 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 56 kDa for AKT pS473. Other band(s): unspecific.
Western Blot: AKT1 [p Ser473] Antibody [NB600-590]

Western Blot: AKT1 [p Ser473] Antibody [NB600-590]

AKT1-[p-Ser473]-Antibody-Western-Blot-NB600-590-img0012.jpg
Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry of AKT1 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 100 dilution for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.
Immunohistochemistry: AKT1 [p Ser473] Antibody [NB600-590] - Immunohistochemistry at higher magnification of AKT1 [p Ser473] antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required.Primary antibody: AKT1 pS473 antibody at 100 dilution for 1 h at RT.Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT.Localization: AKT1 pS473 is nuclear and occasionally cytoplasmic.
Dot Blot: AKT1 [p Ser473] Antibody [NB600-590]
AKT1 [p Ser473] Antibody

AKT1 [p Ser473] Antibody

Dot Blot of Rabbit Anti-AKT pS473 Antibody. Dilutions in Columns: (1) 100ng, (2) 33.33ng, (3) 11.11ng, (4) 3.7ng, (5) 1.23ng. Tested BSA Peptide Reactivity in Rows: (A) AKT1-BSA, (B) AKT1 pT308-BSA, (C) AKT1 S473-BSA, (D) AKT1 pS473-BSA, (E) CDC27 T244-BSA, (F) CDC27 pT244-BSA, (G) BSA control. Primary Antibody: Anti-AKTpS473 at 1ug/mL overnight at 2-8C. Secondary Antibody: Goat anti-Rabbit IgG HRP
AKT1 [p Ser473] Antibody

AKT1 [p Ser473] Antibody

Immunohistochemistry of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.
AKT1 [p Ser473] Antibody

AKT1 [p Ser473] Antibody

Immunohistochemistry at higher magnification of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10,000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.
AKT1 [p Ser473] Antibody

AKT1 [p Ser473] Antibody

Western Blot of Rabbit anti-AKT pS473 antibody. Lane 1: AKT1 Recombinant Protein

Applications for AKT1 [p Ser473] Antibody - BSA Free

Application
Recommended Usage

ELISA

1:15000-1:60000

Immunocytochemistry/ Immunofluorescence

1:40

Immunohistochemistry

1:100-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:200-1:1000
Application Notes
This This product is phospho specific for pS473 and is tested in ELISA, western blotting, immunohistochemistry (formalin-fixed paraffin-embedded sections), and immunofluorescence. By immunoblot a single band of the expected apparent molecular weight ~56kDa is observed. For immunohistochemistry no pre-treatment of sample is required.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

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

Format

BSA Free

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 short term. Aliquot and store at -80C 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)

Gene Symbol

AKT1

UniProt

P31749

Additional Akt1 Products

Product Documents for AKT1 [p Ser473] Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for AKT1 [p Ser473] Antibody - 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 - 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 - 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
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