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    pi-3-kinase-p85-alpha-antibody-epr5513-ab133595.pdf

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Signal Transduction Signaling Pathway Lipid Signaling Lipid Kinases
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
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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

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Primary
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Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Primary
Product image
Anti-PI 3 Kinase p85 alpha antibody [SP62] - BSA and Azide free (ab236440)

View more associated products

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

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 5.50 x 10 -12 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5513
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Lipid Signaling
    • Lipid Kinases
    • Cancer
    • Signal transduction
    • Other
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-PI 3 Kinase p85 alpha antibody [EPR5513] - BSA and Azide free (ab240072)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell line (ab265116)
  • KO cell lysates

    • Human PIK3R1 (PI 3 Kinase p85 alpha) knockout HeLa cell lysate (ab257029)
  • Positive Controls

    • Raji whole cell lysate (ab30124)
    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

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

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.

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/IF,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.
  • 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 [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)

Protocols

  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (14)

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

ab133595 has been referenced in 14 publications.

  • Liu G  et al. ARHGAP20 Expression Inhibited HCC Progression by Regulating the PI3K-AKT Signaling Pathway. J Hepatocell Carcinoma 8:271-284 (2021). PubMed: 33907697
  • 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
  • Wang W  et al. Girdin interaction with vimentin induces EMT and promotes the growth and metastasis of pancreatic ductal adenocarcinoma. Oncol Rep 44:637-649 (2020). PubMed: 32467989
  • Liu B  et al. Kuijieyuan Decoction Improved Intestinal Barrier Injury of Ulcerative Colitis by Affecting TLR4-Dependent PI3K/AKT/NF-?B Oxidative and Inflammatory Signaling and Gut Microbiota. Front Pharmacol 11:1036 (2020). PubMed: 32848725
  • Kang LH  et al. Ganoderic acid A protects lens epithelial cells from UVB irradiation and delays lens opacity. Chin J Nat Med 18:934-940 (2020). PubMed: 33357724
View all Publications for this product

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