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-Paraffin, Immunohistochemistry-Frozen, Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 104A282

Format

BSA Free
View all Akt1 Primary 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.
Immunohistochemistry-Frozen: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - Rat mammary carcinoma tissue section stained with AKT1 [p Ser473] Antibody (104A282). IHC-Fr image submitted by a verified customer review.
Immunohistochemistry-Paraffin: AKT1 [p Ser473] Antibody (104A282) [NB100-56749] - IHC analysis of an FFPE human breast carcinoma tissue section using 1:250 dilution of pSer473 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. Some cancer cells depicted nuclear stianing also. Staining was performed by Histowiz.
AKT1 [p Ser473] Antibody (104A282) - BSA Free

Western Blot: AKT1 [p Ser473] Antibody (104A282) - BSA Free [NB100-56749] -

SP1/PTEN mediates lung infection in mice and the activity of the Akt signaling pathway. A, survival days of the mice after si-SP1 and si-PTEN administration (p < 0.01, the Kaplan-Meier analysis); B, bacterial load in murine lung tissues examined by CFU analysis (*p < 0.05, the one-way ANOVA); C, infiltration of inflammatory cells in murine lung tissues examined by HE staining (*p < 0.05, the one-way ANOVA); D, epithelial cell apoptosis in the murine lung tissues after Mtb infection examined by the TUNEL assay (*p < 0.05, the one-way ANOVA); E, fibrosis in murine lung tissues determined by Masson’s trichrome staining (*p < 0.05, the one-way ANOVA); F-G, protein levels of SP1 and PTEN and the Akt phosphorylation in murine lung tissues determined by western blot analysis (*p < 0.05, the one-way ANOVA). For animal studies, n = 6 in each group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35420971), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AKT1 [p Ser473] Antibody (104A282) - BSA Free

Western Blot: AKT1 [p Ser473] Antibody (104A282) - BSA Free [NB100-56749] -

EZH2/HMGA2 regulates the PI3K/AKT signaling. MI rats were subjected to injection of EZH2-OE or EZH2-NC (MI + MSC-EXO + EZH2 NC/EZH2 OE) and EZH2-OE + HMGA2-OE or EZH2-OE + HMGA2-NC. A HMGA2 protein expression in myocardial tissues in response to EZH2-OE + HMGA2-OE examined using western blot (Additional file 5: Fig. S5). B Weight change in rats after EZH2-OE + HMGA2-OE treatment. C Changes in HMI in rats after EZH2-OE + HMGA2-OE treatment. D Measurement of Dd, Sd, and LVEF by echocardiography at 2 weeks after LAD in rats. E Measurement of LVMI by echocardiography in rats. F KEGG pathway analysis of gene enrichment pathways [29]. G Changes in PI3K/AKT pathway in rat myocardium examined using western blot (Additional file 6: Fig. S6). All data are expressed as mean +/- SD (n = 6/group, *p < 0.05 vs. Sham, DMSO, MI + MSC-EXO + EZH2-NC or EZH2-OE + HMGA2-NC group determined by unpaired t test or one-way ANOVA) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35264108), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
AKT1 [p Ser473] Antibody (104A282) - BSA Free

Western Blot: AKT1 [p Ser473] Antibody (104A282) - BSA Free [NB100-56749] -

SP1/PTEN mediates lung infection in mice and the activity of the Akt signaling pathway. A, survival days of the mice after si-SP1 and si-PTEN administration (p < 0.01, the Kaplan-Meier analysis); B, bacterial load in murine lung tissues examined by CFU analysis (*p < 0.05, the one-way ANOVA); C, infiltration of inflammatory cells in murine lung tissues examined by HE staining (*p < 0.05, the one-way ANOVA); D, epithelial cell apoptosis in the murine lung tissues after Mtb infection examined by the TUNEL assay (*p < 0.05, the one-way ANOVA); E, fibrosis in murine lung tissues determined by Masson’s trichrome staining (*p < 0.05, the one-way ANOVA); F-G, protein levels of SP1 and PTEN and the Akt phosphorylation in murine lung tissues determined by western blot analysis (*p < 0.05, the one-way ANOVA). For animal studies, n = 6 in each group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35420971), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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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.

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

Customer Reviews for AKT1 [p Ser473] Antibody (104A282) - BSA Free (2)

5 out of 5
2 Customer Ratings
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Showing  1 - 2 of 2 reviews Showing All
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  • AKT1 [p Ser473] Antibody (104A282)
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mammary carcinoma
    Species: Rat
    Verified Customer | Posted 08/10/2021
    Mammary carcinoma
    AKT1 [p Ser473] Antibody (104A282) - BSA Free NB100-56749
    Bio-Techne Response
    This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.
  • Name: Liyong Zhang
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 08/16/2014
    A review for p-S473-AKT antibody (NB100-56749)
    AKT1 [p Ser473] Antibody (104A282) - BSA Free NB100-56749

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Protocols

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FAQs for AKT1 [p Ser473] Antibody (104A282) - 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.

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