Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

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
  • Tested applications
    Suitable for: WB, ICC, IHC-P, ICC/IF, ELISAmore 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 cell lysate. 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.

Properties

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).
ICC 1/100.
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use at an assay dependent concentration.
ELISA 1/2000.

Only tested in Peptide ELISA

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.
  • Information by UniProt
  • Database links
  • 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 subunit alpha antibody
    • PI3-kinase regulatory subunit alpha antibody
    • PI3-kinase subunit p85-alpha antibody
    • PI3K 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

Anti-PI 3 Kinase p85 alpha antibody [M253] images

  • 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

    Lane 1 : A431 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : A431 cell lysate


    Predicted band size : 85 kDa
    Observed band size : 85 kDa
  • 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.

  • 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.

  • 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.

References for Anti-PI 3 Kinase p85 alpha antibody [M253] (ab86714)

This product has been referenced in:
  • Tantai JC  et al. Heterophyllin B inhibits the adhesion and invasion of ECA-109 human esophageal carcinoma cells by targeting PI3K/AKT/ß-catenin signaling. Mol Med Rep 13:1097-104 (2016). WB ; Human . Read more (PubMed: 26647768) »
  • Qian J  et al. Downregulation of HIF-1a inhibits the proliferation and invasion of non-small cell lung cancer NCI-H157 cells. Oncol Lett 11:1738-1744 (2016). Read more (PubMed: 26998070) »

See all 9 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Julie Kim

Verified customer

Submitted Apr 17 2017

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
Username

Dr. Armen Petrosyan

Verified customer

Submitted Oct 21 2015

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
Username

Abcam user community

Verified customer

Submitted Oct 21 2015

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
Username

Abcam user community

Verified customer

Submitted Oct 19 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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