Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

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Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
  • OI-RD Scanning - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
  • Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PI 3 Kinase p85 alpha antibody [EPR5513]
    See all PI 3 Kinase p85 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to a region within the N terminus of Human PI 3 Kinase p85 alpha (UniProt ID: P27986)

  • Positive control

    • HeLa, Jurkat, Raji, K562, MCF7, C6, Raw 264.7, PC12, NIH3T3 cell lysates

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133595 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
WB
Human
Application Abreviews Notes
WB
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 83 kDa).
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 83 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC,IHC-P or IP.

    • 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

      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 [EPR5513] (ab133595)
      Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PI 3 Kinase p85 alpha knockout HAP1 cell lysate (20 µg)
      Lane 3: Jurkat cell lysate (20 µg)
      Lane 4: THP1 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab133595 was shown to specifically react with PI 3 Kinase p85 alpha when PI 3 Kinase p85 alpha knockout samples were used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. ab133595 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : PIK3R1 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 83 kDa
      Observed band size: 90 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab133595 was shown to react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type HeLa and PIK3R1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : Raji cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : C6 cell lysate
      Lane 6 : Raw 264.7 cell lysate
      Lane 7 : PC12 cell lysate
      Lane 8 : NIH3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution

      Predicted band size: 83 kDa

    • OI-RD Scanning - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      OI-RD Scanning - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
      Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

    References (10)

    Publishing research using ab133595? Please let us know so that we can cite the reference in this datasheet.

    ab133595 has been referenced in 10 publications.

    • Li Y  et al. Protective effect of CTRP6 on cerebral ischemia/reperfusion injury by attenuating inflammation, oxidative stress and apoptosis in PC12 cells. Mol Med Rep 22:344-352 (2020). PubMed: 32377750
    • Molinaro A  et al. Insulin-Driven PI3K-AKT Signaling in the Hepatocyte Is Mediated by Redundant PI3Ka and PI3Kß Activities and Is Promoted by RAS. Cell Metab 29:1400-1409.e5 (2019). PubMed: 30982732
    • Du J  et al. Astragaloside IV protects cardiomyocytes from hypoxia-induced injury by down-regulation of lncRNA GAS5. Biomed Pharmacother 116:109028 (2019). PubMed: 31136949
    • Fu Y  et al. Declination of long noncoding RNA paternally expressed gene 10 inhibits A375 cells proliferation, migration, and invasion via mediating microRNA-33a. J Cell Biochem 120:19868-19877 (2019). PubMed: 31318088
    • Yang Q  et al. Lidocaine alleviates cytotoxicity-resistance in lung cancer A549/DDP cells via down-regulation of miR-21. Mol Cell Biochem 456:63-72 (2019). PubMed: 30644017
    • Xiao X  et al. The better effects of microbubble ultrasound transfection of miR-940 on cell proliferation inhibition and apoptosis promotion in human cervical cancer cells. Onco Targets Ther 12:6813-6824 (2019). PubMed: 31686839
    • Zhao J  et al. Salidroside mitigates hydrogen peroxide-induced injury by enhancement of microRNA-27a in human trabecular meshwork cells. Artif Cells Nanomed Biotechnol 47:1758-1765 (2019). PubMed: 31062616
    • Zhu B  et al. Downregulation of Krüppel-like factor 1 inhibits the metastasis and invasion of cervical cancer cells. Mol Med Rep 18:3932-3940 (2018). PubMed: 30132534
    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
    • Pan H & Zhu L Angelica sinensis polysaccharide protects rat cardiomyocytes H9c2 from hypoxia-induced injury by down-regulation of microRNA-22. Biomed Pharmacother 106:225-231 (2018). PubMed: 29960169

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