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Signal Transduction Signaling Pathway Lipid Signaling Lipid Kinases
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Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

  • Datasheet
  • SDS
Reviews (7) Submit a question References (117)

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Abpromise

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Western blot - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

Key features and details

  • Mouse monoclonal [M253] to PI 3 Kinase p85 alpha
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG2a

You may also be interested in

Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Protein
Product image
Recombinant Human PI 3 Kinase p85 alpha protein (ab84769)

View more associated products

Overview

  • Product name

    Anti-PI 3 Kinase p85 alpha antibody [M253]
    See all PI 3 Kinase p85 alpha primary antibodies
  • Description

    Mouse monoclonal [M253] to PI 3 Kinase p85 alpha
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken
  • Immunogen

    Recombinant fragment corresponding to Human PI 3 Kinase p85 alpha (C terminal).

  • Positive control

    • A431 or Jurkat whole cell lysate (ab7899). This antibody gave a positive result in IHC in the following FFPE tissue: Human skeletal muscle.
  • General notes

    Phosphoinositide 3-kinase (PI3K) phosphorylates phosphatidylinositol (PI), PI-4-phosphate and PI-4,5-bisphosphate to catalyze the production of PI-3,4,5-triphosphate. Growth factors and hormones activate PI 3 Kinase to coordinate various cellular events, such as cell growth, cell cycle entry, cell migration and cell survival.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 50% Glycerol, 49% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    M253
  • Isotype

    IgG2a
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Lipid Signaling
    • Lipid Kinases
    • Cancer
    • Signal transduction
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Signal transducers
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Obesity

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) (ab96871)
  • Isotype control

    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)
    • A431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Human PI 3 Kinase p85 alpha protein (ab84769)
  • Related Products

    • 100x Citrate Buffer pH 6.0 (ab64236)
    • Normal Goat Serum (ab7481)

Applications

Our Abpromise guarantee covers the use of ab86714 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Necessary for the insulin-stimulated increase in glucose uptake and glycogen synthesis in insulin-sensitive tissues.
  • Tissue specificity

    Isoform 2 is expressed in skeletal muscle and brain, and at lower levels in kidney and cardiac muscle. Isoform 2 and isoform 4 are present in skeletal muscle (at protein level).
  • Sequence similarities

    Belongs to the PI3K p85 subunit family.
    Contains 1 Rho-GAP domain.
    Contains 2 SH2 domains.
    Contains 1 SH3 domain.
  • Domain

    The SH3 domain mediates the binding to CBLB, and to HIV-1 Nef.
  • Post-translational
    modifications

    Polyubiquitinated in T-cells by CBLB; which does not promote proteasomal degradation but impairs association with CD28 and CD3Z upon T-cell activation.
    Phosphorylated. Dephosphorylated by PTPRJ.
  • Target information above from: UniProt accession P27986 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5295 Human
    • Entrez Gene: 18708 Mouse
    • Entrez Gene: 25513 Rat
    • Omim: 171833 Human
    • SwissProt: P27986 Human
    • SwissProt: P26450 Mouse
    • SwissProt: Q63787 Rat
    • Unigene: 132225 Human
    • Unigene: 604502 Human
    • Unigene: 259333 Mouse
    • Unigene: 10599 Rat
    see all
  • Alternative names

    • GRB1 antibody
    • p85 alpha antibody
    • p85 antibody
    • P85A_HUMAN antibody
    • Phosphatidylinositol 3 kinase associated p 85 alpha antibody
    • Phosphatidylinositol 3 kinase regulatory 1 antibody
    • Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 1 (p85 alpha) antibody
    • Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha antibody
    • Phosphatidylinositol 3-kinase regulatory subunit alpha antibody
    • Phosphoinositide 3 kinase, regulatory subunit 1 (alpha) antibody
    • PI3 kinase p85 antibody
    • PI3 kinase p85 subunit alpha antibody
    • PI3-kinase regulatory subunit alpha antibody
    • PI3-kinase subunit p85-alpha antibody
    • PI3K antibody
    • PI3K p85 antibody
    • PI3K regulatory subunit alpha antibody
    • Pik3r1 antibody
    • PtdIns 3 kinase p85 alpha antibody
    • PtdIns-3-kinase regulatory subunit alpha antibody
    • PtdIns-3-kinase regulatory subunit p85-alpha antibody
    see all

Images

  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
    Lane 1 : Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714) at 1/1000 dilution
    Lane 2 : Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714) at 1/2000 dilution
    Lane 3 : Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714) at 1/4000 dilution

    All lanes : A431 cell lysate

    Predicted band size: 85 kDa
    Observed band size: 85 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

    Immunocytochemical labeling of PI 3 Kinase p85 in aldehyde-fixed and NP-40-permeabilized A431 cells. The cells were labeled with ab86714, then the antibody was detected using appropriate secondary antibody conjugated to DyLight® 594.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

    IHC image of PI 3 Kinase p85 staining in Human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (ab64236) (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab86714, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

    ICC/IF image of ab86714 stained SKNSH cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86714, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) (ab96871) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (117)

    Publishing research using ab86714? Please let us know so that we can cite the reference in this datasheet.

    ab86714 has been referenced in 117 publications.

    • Yan J  et al. Integrative transcriptomic and proteomic analysis reveals CD9/ITGA4/PI3K-Akt axis mediates trabecular meshwork cell apoptosis in human glaucoma. J Cell Mol Med 24:814-829 (2020). PubMed: 31680442
    • Yang HZ  et al. Resveratrol inhibits the proliferation of melanoma cells by modulating cell cycle. Int J Food Sci Nutr 71:84-93 (2020). PubMed: 31154861
    • Zhang Y  et al. Icariin protects against sodium azide-induced neurotoxicity by activating the PI3K/Akt/GSK-3ß signaling pathway. PeerJ 8:e8955 (2020). PubMed: 32341897
    • Piao X  et al. Picroside II Improves Severe Acute Pancreatitis-Induced Intestinal Barrier Injury by Inactivating Oxidative and Inflammatory TLR4-Dependent PI3K/AKT/NF-?B Signaling and Improving Gut Microbiota. Oxid Med Cell Longev 2020:3589497 (2020). PubMed: 32351672
    • Bao J  et al. Rapamycin protects chondrocytes against IL-18-induced apoptosis and ameliorates rat osteoarthritis. Aging (Albany NY) 12:5152-5167 (2020). PubMed: 32182210
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-7 of 7 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (RM-1 - mouse prostate cancer cells)
    Permeabilization
    Yes - 0.2% Triton X-100 in PBS
    Specification
    RM-1 - mouse prostate cancer cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Dr. Armen Petrosyan

    Verified customer

    Submitted Dec 07 2020

    Immunocytochemistry abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (Melanoma cells)
    Permeabilization
    No
    Specification
    Melanoma cells
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 01 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Melanoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA
    Permeabilization
    Yes - PBS-Tween
    Specification
    Melanoma
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 07 2020

    Western blot abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Bos taurus Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient gel)
    Loading amount
    20 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Julie Kim

    Verified customer

    Submitted Apr 17 2017

    Western blot abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (RWPE - prostate cells)
    Gel Running Conditions
    Reduced Denaturing (10% SDS-PAGE)
    Loading amount
    30 µg
    Specification
    RWPE - prostate cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Dr. Armen Petrosyan

    Verified customer

    Submitted Oct 21 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (LNCaP - prostate cancer cells)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    LNCaP - prostate cancer cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 21 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-PI 3 Kinase p85 alpha antibody [M253]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Rat prostate)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    Rat prostate
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 19 2015

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