Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] (ab40776)

Overview

  • Product nameAnti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y]
    See all PI 3 Kinase catalytic subunit alpha primary antibodies
  • Description
    Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha
  • Tested applicationsSuitable for: Flow Cyt, WB, ICC/IF, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PI 3 Kinase catalytic subunit alpha aa 1000 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: Jurkat, MCF-7, Raw264.7 and NIH/3T3 cell lysates. ICC/IF: HeLa and Jurkat cells. IP: Jurkat cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:

    Anti-PI 3 Kinase catalytic subunit alpha antibody (Alexa Fluor® 488) [EP383Y] (ab202671)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab40776 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
Flow Cyt Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).
ICC/IF 1/100 - 1/250.
IP 1/20 - 1/30.
  • Application notesIs unsuitable for IHC-P.
  • Target

    • FunctionPhosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2.
    • Involvement in diseaseDefects in PIK3CA are associated with colorectal cancer (CRC) [MIM:114500].
      Defects in PIK3CA are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
      Defects in PIK3CA are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
      Defects in PIK3CA may underlie hepatocellular carcinoma (HCC) [MIM:114550].
      Defects in PIK3CA are a cause of keratosis seborrheic (KERSEB) [MIM:182000]. A common benign skin tumor. Seborrheic keratoses usually begin with the appearance of one or more sharply defined, light brown, flat macules. The lesions may be sparse or numerous. As they initially grow, they develop a velvety to finely verrucous surface, followed by an uneven warty surface with multiple plugged follicles and a dull or lackluster appearance.
    • Sequence similaritiesBelongs to the PI3/PI4-kinase family.
      Contains 1 C2 domain.
      Contains 1 PI3K/PI4K domain.
    • Information by UniProt
    • Database links
    • Alternative names
      • 5-bisphosphate 3-kinase 110 kDa catalytic subunit alpha antibody
      • 5-bisphosphate 3-kinase catalytic subunit alpha isoform antibody
      • caPI3K antibody
      • CLOVE antibody
      • CWS5 antibody
      • MCAP antibody
      • MCM antibody
      • MCMTC antibody
      • MGC142161 antibody
      • MGC142163 antibody
      • p110 alpha antibody
      • p110alpha antibody
      • Phosphatidylinositol 3 kinase catalytic alpha polypeptide antibody
      • Phosphatidylinositol 3 kinase catalytic 110 KD alpha antibody
      • Phosphatidylinositol 4 5 bisphosphate 3 kinase catalytic subunit alpha antibody
      • Phosphatidylinositol 4 5 bisphosphate 3 kinase catalytic subunit alpha isoform antibody
      • Phosphatidylinositol 4,5 bisphosphate 3 kinase 110 kDa catalytic subunit alpha antibody
      • Phosphatidylinositol-4 antibody
      • Phosphoinositide 3 kinase catalytic alpha polypeptide antibody
      • PI3 kinase p110 subunit alpha antibody
      • PI3-kinase subunit alpha antibody
      • PI3K antibody
      • PI3K-alpha antibody
      • PI3KC A antibody
      • PIK3C A antibody
      • Pik3ca antibody
      • PK3CA antibody
      • PK3CA_HUMAN antibody
      • PtdIns 3 kinase p110 antibody
      • PtdIns-3-kinase subunit alpha antibody
      • PtdIns-3-kinase subunit p110-alpha antibody
      • Serine/threonine protein kinase PIK3CA antibody
      see all

    Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] images

    • ab40776 staining PI 3 Kinase catalytic subunit alpha in the human cell line Jurkat (human acute T cell leukemia) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)



    • Predicted band size : 110 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PI 3 Kinase catalytic subunit alpha knockout HAP1 cell lysate (20 µg)
      Lane 3: Jurkat cell lysate (20 µg)
      Lane 4: MCF7 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab40776 observed at 100,120 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab40776 was shown to recognize PI 3 Kinase catalytic subunit alpha when PI 3 Kinase catalytic subunit alpha knockout samples were used, along with additional cross-reactive bands. Wild-type and PI 3 Kinase catalytic subunit alpha knockout samples were subjected to SDS-PAGE. ab40776 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776
      secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] (ab40776) at 1/5000 dilution (purified)

      Lane 1 : Jurkat whole cell lysate
      Lane 2 : MCF-7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 110 kDa
      Observed band size : 110 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • All lanes : Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] (ab40776) at 1/5000 dilution (purified)

      Lane 1 : Raw264.7 whole cell lysate
      Lane 2 : NIH/3T3 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 110 kDa
      Observed band size : 110 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling PI 3 Kinase catalytic subunit alpha with purified ab40776 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • ab40776 (purified) at a dilution of 1/20 immunoprecipitating PI 3 Kinase catalytic subunit alpha in Jurkat whole cell lysate.

      Lane 1 (input): Jurkat whole cell lysate (10µg)

      Lane 2 (+): ab40776 + Jurkat whole cell lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40776 in Jurkat whole cell lysate.

      For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] (ab40776) at 1/5000 dilution (unpurified) + Jurkat cell lysate at 10 µg

      Predicted band size : 110 kDa
      Observed band size : 110 kDa
    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PI 3 Kinase catalytic subunit alpha with unpurified ab40776 at a dilution of 1/100.

    References for Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y] (ab40776)

    This product has been referenced in:
    • Hennessy BT  et al. A Technical Assessment of the Utility of Reverse Phase Protein Arrays for the Study of the Functional Proteome in Non-microdissected Human Breast Cancers. Clin Proteomics 6:129-51 (2010). Dot blot ; Human . Read more (PubMed: 21691416) »
    • Levin VA  et al. Different changes in protein and phosphoprotein levels result from serum starvation of high-grade glioma and adenocarcinoma cell lines. J Proteome Res 9:179-91 (2010). Read more (PubMed: 19894763) »

    See all 5 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing
    Sample Zebrafish Tissue lysate - whole (24hpf embryos)
    Specification 24hpf embryos
    Blocking step Odyssey blocking buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 24°C
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    Submitted Mar 28 2014

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