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    pi-3-kinase-catalytic-subunit-alphapik3ca-antibody-ep383y-ab40776.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 catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

  • Datasheet
  • SDS
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Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Flow Cytometry (Intracellular) - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
  • Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha/PIK3CA
  • Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y]
    See all PI 3 Kinase catalytic subunit alpha/PIK3CA primary antibodies
  • Description

    Rabbit monoclonal [EP383Y] to PI 3 Kinase catalytic subunit alpha/PIK3CA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human PI 3 Kinase catalytic subunit alpha/PIK3CA aa 1000 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P42336

  • Positive control

    • WB: Jurkat, MCF-7, Raw264.7 and NIH/3T3 cell lysates. ICC/IF: HeLa and Jurkat cells. IP: Jurkat whole cell lysate (ab7899).
  • 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.

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP383Y
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)
    • Alexa Fluor® 488 Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab202671)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • MCF7 whole cell lysate (ab29537)
    • Jurkat whole cell lysate (ab30128)
    • 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 catalytic subunit alpha/PIK3CA protein (ab159127)

Applications

The Abpromise guarantee

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
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.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
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.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Phosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2.
  • Involvement in disease

    Defects 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 similarities

    Belongs to the PI3/PI4-kinase family.
    Contains 1 C2 domain.
    Contains 1 PI3K/PI4K domain.
  • Target information above from: UniProt accession P42336 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5290 Human
    • Entrez Gene: 18706 Mouse
    • Entrez Gene: 170911 Rat
    • Omim: 171834 Human
    • SwissProt: P42336 Human
    • SwissProt: P42337 Mouse
    • Unigene: 553498 Human
    • Unigene: 85701 Human
    • Unigene: 260521 Mouse
    see all
  • 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
    • PI 3 Kinase catalytic subunit alpha 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

Images

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PI 3 Kinase catalytic subunit alpha/PIK3CA 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 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab40776 was shown to recognize PI 3 Kinase catalytic subunit alpha/PIK3CA  when PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were used, along with additional cross-reactive bands. Wild-type and PI 3 Kinase catalytic subunit alpha/PIK3CA 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Flow Cytometry (Intracellular) - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

    ab40776 staining PI 3 Kinase catalytic subunit alpha/PIK3CA in the human cell line Jurkat (human acute T cell leukemia) by intracellular 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) 

     

     

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776) at 1/1000 dilution

    Lane 1 : Jurkat Whole Cell Lysate
    Lane 2 : Mouse Brain Tissue Lysate
    Lane 3 : Rat Brain Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 110 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    Lanes 1 - 3: Merged signal (red and green). Green - ab40776 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab40776 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA 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
    All lanes : 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
  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA 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
    All lanes : 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 - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA 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).

  • Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

    ab40776 (purified) at a dilution of 1/20 immunoprecipitating PI 3 Kinase catalytic subunit alpha/PIK3CA 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 for detection (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA 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 - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with unpurified ab40776 at a dilution of 1/100.

  • Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)
    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (ab40776)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (56)

Publishing research using ab40776? Please let us know so that we can cite the reference in this datasheet.

ab40776 has been referenced in 56 publications.

  • Chen Q  et al. Silencing of PSMC2 inhibits development and metastasis of prostate cancer through regulating proliferation, apoptosis and migration. Cancer Cell Int 21:235 (2021). PubMed: 33902600
  • Huang Y  et al. GSG2 Promotes Development and Predicts Poor Prognosis of Ovarian Cancer. Cancer Manag Res 13:499-508 (2021). PubMed: 33500663
  • Wang X & Zhu Y Circ_0000020 elevates the expression of PIK3CA and facilitates the malignant phenotypes of glioma cells via targeting miR-142-5p. Cancer Cell Int 21:79 (2021). PubMed: 33509213
  • Tang D  et al. Silencing LMNB1 Contributes to the Suppression of Lung Adenocarcinoma Development. Cancer Manag Res 13:2633-2642 (2021). PubMed: 33776481
  • Feng Z & Yu CH PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking and invasive cell migration. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 33947811
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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Western blot abreview for Anti-PI 3 Kinase catalytic subunit alpha antibody [EP383Y]

Inconclusive
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Mar 28 2014

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