PI 3-Kinase p85 alpha Antibody (A3-D0-R)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32817

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32817
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Key Product Details

Species Reactivity

Validated:

Human

Applications

Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG1 Clone # A3-D0-R

Concentration

1 mg/ml

View all PI 3-Kinase p85 alpha Antibodies »

Product Summary for PI 3-Kinase p85 alpha Antibody (A3-D0-R)

Immunogen

Recombinant protein within Human PI3-kinase p85 subunit alpha aa 19-219 / 724. (Uniprot: P27986)

Localization

Cytosol, Membrane, Nucleus.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

84 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 PI 3-Kinase p85 alpha Antibody (A3-D0-R)

PI 3-Kinase p85 alpha Antibody (A3-D0-R)

Immunocytochemistry/ Immunofluorescence: PI 3-Kinase p85 alpha Antibody (A3-D0-R) [NBP3-32817]

Immunocytochemistry/ Immunofluorescence: PI 3-Kinase p85 alpha Antibody (A3-D0-R) [NBP3-32817] - Immunocytochemistry analysis of U-937 cells labeling PI 3-Kinase p85 alpha with Mouse anti-PI 3-Kinase p85 alpha antibody (NBP3-32817) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PI 3-Kinase p85 alpha antibody (NBP3-32817) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.
PI 3-Kinase p85 alpha Antibody (A3-D0-R)

Western Blot: PI 3-Kinase p85 alpha Antibody (A3-D0-R) [NBP3-32817]

Western Blot: PI 3-Kinase p85 alpha Antibody (A3-D0-R) [NBP3-32817] - Western blot analysis of PI 3-Kinase p85 alpha on different lysates with Mouse anti-PI 3-Kinase p85 alpha antibody (NBP3-32817) at 1/5,000 dilution.
Lane 1: SW480 cell lysate
Lane 2: A431 cell lysate
Lane 3: 293T cell lysate
Lane 4: Jurkat cell lysate
Lane 5: Raji cell lysate
Lane 6: MCF7 cell lysate
Lane 7: A549 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 84 kDa
Observed band size: 84 kDa

Exposure time: 1 minute 40 seconds;

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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Applications for PI 3-Kinase p85 alpha Antibody (A3-D0-R)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Western Blot

1:1000-1:5000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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: PI 3-Kinase p85 alpha

Phosphatidylinositol 3-kinase (PI3K) p85 alpha is a regulatory subunit that heterodimerizes with a catalytic subunit to form a Class IA PI3K enzyme complex, which plays an important role in the immune system (1,2). The P13K pathway is involved in many diverse processes including growth, metabolism, proliferation, and survival (2). PI3Ks are typically activated by cytokine receptors and are responsible for phosphorylation of the 3'-hydroxyl group of PI and its derivatives (3). p85 alpha is one of five regulatory subunit proteins which also includes p55 alpha, p50 alpha, p85 beta, and p55 gamma, and can bind to two of the three catalytic subunits (p110 alpha or p110 delta) (1,2). p85 alpha, p55 alpha, and p50 alpha proteins are all synthesized by the same PIK3R1 gene by alternative splicing (1). Structurally, PI 3-Kinase p85 alpha contains SRC homology 3 (SH3 domain), followed by a Bcr homology (BH) domain flanked by two proline-rich regions, then a N-terminal SH2 domain, an inter-SH2 domain, and C-terminal SH2 domain (1,2). PI 3-Kinase p85 alpha protein consists of 724 amino acids (aa) in length with a theoretical molecular weight of 83.5 kDa (2,4). The primary role for the p85 subunit is interaction with cell surface receptors and acts as an adapter for the stabilization and recruitment of the p110 catalytic subunit to the plasma membrane (1,3). Additionally, p85 has been shown to function in both interleukin-2 receptor (IL2R) and erythropoietin receptor (EpoR) endocytosis (3).

The PI3K pathway functions in a broad range of cellular processes, so it is understandable that pathway dysfunction can lead to an array of diseases and disorders (2,5). Elevated PI3K signaling is a key feature of many cancers (5). PI3K pathway dysregulation has also been implicated in neurological, metabolic, and cardiovascular disorders (5). Furthermore, both overactivation or under-activation of the PI3K delta (p85 alpha subunit + p110 delta subunit) pathway has been shown to cause immunodeficiency and pathologies related to immune system dysfunction (2). Therapeutics to target the PI3K pathway and treat related cancers include PI3K inhibitors and, specifically, isoform-selective inhibitors which have a lot of promise when used as part of a combination therapy (5).

References

1. Okkenhaug, K., & Vanhaesebroeck, B. (2001). New responsibilities for the PI3K regulatory subunit p85 alpha. Science's STKE : signal transduction knowledge environment. https://doi.org/10.1126/stke.2001.65.pe1

2. Nunes-Santos, C. J., Uzel, G., & Rosenzweig, S. D. (2019). PI3K pathway defects leading to immunodeficiency and immune dysregulation. The Journal of allergy and clinical immunology. https://doi.org/10.1016/j.jaci.2019.03.017

3. Chen, P. H., Yao, H., & Huang, L. J. (2017). Cytokine Receptor Endocytosis: New Kinase Activity-Dependent and -Independent Roles of PI3K. Frontiers in endocrinology. https://doi.org/10.3389/fendo.2017.00078

4. Uniprot (P27986)

5. Fruman, D. A., Chiu, H., Hopkins, B. D., Bagrodia, S., Cantley, L. C., & Abraham, R. T. (2017). The PI3K Pathway in Human Disease. Cell. https://doi.org/10.1016/j.cell.2017.07.029

Long Name

PI 3-Kinase p85 alpha

Alternate Names

GRB1, PI 3Kinase p85 alpha, PIK3R1

Gene Symbol

PIK3R1

Additional PI 3-Kinase p85 alpha Products

Product Documents for PI 3-Kinase p85 alpha Antibody (A3-D0-R)

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Certificate of Analysis

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Product Specific Notices for PI 3-Kinase p85 alpha Antibody (A3-D0-R)

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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