• Product NamePI 3 Kinase p85 alpha Rabbit mAb
• Clone No.SU04-07
• PurificationProA affinity purified
• ApplicationsWB, ICC/IF, IHC, FC
• Species ReactivityHu, Ms, Rt
• Immunogen DescriptionSynthetic peptide within C-terminal human PI 3 Kinase p85 alpha.
• 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
  
• Accession No. Swiss-Prot#:P27986
• UniprotP27986
• Gene ID5295;
• Calculated MW 84 kDa
• Concentration 1 mg/mL
• Formulation 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
• Storage Store at -20˚C

• WB: 1:1,000-1:2,000
  IHC: 1:50-1:200
  ICC: 1:50-1:200
  
  FC: 1:50-1:100

Phosphatidylinositol 3-kinase (PI 3-kinase) phosphorylates the 3' OH position of the inositol ring of inositol lipids and is composed of p85 and p110 subunits. PI 3-kinase p85 lacks PI 3-kinase activity and acts as an adapter, coupling p110 to activated protein tyrosine kinase. Two forms of p85 have been described (p85α and p85β), each possessing one SH3 and two SH2 domains. PI 3-kinase p85α, also known as GRB1, phosphatidylinositol 3-kinase regulatory 1 or p85, is a 724 amino acid protein that exists as four alternatively spliced isoforms. Involved in insulin metabolism, defects in the PI 3-kinase p85α gene have been linked to insulin resistance. PI 3-kinase p85α is polyubiquitinated in T-cells by Cbl-b, and has multiple phosphorylated amino acid residues, including a phosphorylated tyrosine residue at position 467.

• Western blot analysis of PI 3 Kinase p85 alpha on different lysates using anti-PI 3 Kinase p85 alpha antibody at 1/1,000 dilution. Positive control:
  Lane 1: MCF-7
  Lane 2: Raji
• Flow cytometric analysis of HepG2 cells with PI 3 Kinase p85 alpha antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody
• Figure legend: Western blot showing PIK3R1 (PI 3 Kinase p85 alpha) protein expression in wild-type (WT) and PIK3R1 shRNA knockdown human ACHN cells. GAPDH served as the loading control.Conclusion: Since this antibody detects endogenous PIK3R1 protein in WT but not in shRNA knockdown cell lysate, this antibody is highly specific.
• ICC staining PI 3 Kinase p85 alpha in Hela Cells(red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
• ICC staining PI 3 Kinase p85 alpha in HepG2 cells(red). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
• Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody （1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX
• Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-PI 3 Kinase p85 alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

• Flow Cytometry (FCM) General Protocol
• Immunofluorescence(IC)
• Immunohistochemistry(Polymer)
• Western Blotting