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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)

Reviews (1)Q&A (2)References (15)
Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
  • Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
  • Immunoprecipitation - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
  • Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y384] to PI3 Kinase p110 beta
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-PI3 Kinase p110 beta antibody [Y384]
    See all PI3 Kinase p110 beta primary antibodies

  • Description

    Rabbit monoclonal [Y384] to PI3 Kinase p110 beta

  • Host species

    Rabbit

  • Specificity

    ab32569 recognises PI3-Kinases (PI3-Ks).

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt (Intra),ICC or IHC

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Guinea pig, Cow, Cat, Dog, Pig

  • Immunogen

    Synthetic peptide within Human PI3 Kinase p110 beta. The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat whole cell lysate (ab7899). IP: Jurkat

  • 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.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32569 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)
1/500. Detects a band of approximately 110 kDa (predicted molecular weight: 123 kDa).
IP
1/50.
Notes
WB
1/500. Detects a band of approximately 110 kDa (predicted molecular weight: 123 kDa).
IP
1/50.
Application notes
Is unsuitable for Flow Cyt (Intra),ICC or IHC.

Target

  • Function

    Phosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2.

  • Tissue specificity

    Expressed ubiquitously.

  • Pathway

    Phospholipid metabolism; phosphatidylinositol phosphate biosynthesis.

  • Sequence similarities

    Belongs to the PI3/PI4-kinase family.
    Contains 1 PI3K/PI4K domain.
  • Information by UniProt

  • Database links

    • Alternative names

      • 5-bisphosphate 3-kinase 110 kDa catalytic subunit beta antibody
      • 5-bisphosphate 3-kinase catalytic subunit beta isoform antibody
      • DKFZp779K1237 antibody
      • MGC133043 antibody
      • OTTHUMP00000216901 antibody
      • OTTHUMP00000216904 antibody
      • p110 BETA antibody
      • p110Beta antibody
      • Phosphatidylinositol 3 kinase catalytic beta polypeptide antibody
      • Phosphatidylinositol 4 5 bisphosphate 3 kinase 110 kDa catalytic subunit beta antibody
      • Phosphatidylinositol 4 5 bisphosphate 3 kinase catalytic subunit beta isoform antibody
      • Phosphatidylinositol-4 antibody
      • Phosphoinositide 3 kinase catalytic beta polypeptide antibody
      • PI3 kinase p110 subunit beta antibody
      • PI3-kinase subunit beta antibody
      • PI3K antibody
      • PI3K beta antibody
      • PI3K-beta antibody
      • PI3Kbeta antibody
      • PI3KCB antibody
      • PIK3C1 antibody
      • Pik3cb antibody
      • PK3CB_HUMAN antibody
      • PtdIns 3 kinase p110 antibody
      • PtdIns-3-kinase subunit beta antibody
      • PtdIns-3-kinase subunit p110-beta antibody
      see all

    Images

    • Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
      Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: PI3 Kinase p110 beta knockout HAP1 cell lysate (40 µg)
      Lane 3: Jurkat cell lysate (40 µg)
      Lane 4: K562 cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab32569 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32569 was shown to recognize PI3 Kinase p110 beta when PI3 Kinase p110 beta knockout samples were used, along with additional cross-reactive bands. Wild-type and PI3 Kinase p110 beta knockout samples were subjected to SDS-PAGE. Ab32569 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
      Western blot - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
      Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569) at 1/500 dilution + Jurkat cell lysate

      Predicted band size: 123 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
      Immunoprecipitation - Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)

      PI3 Kinase p110 beta was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 µg with ab32569 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 µg

      Lane 2: ab32569 IP in Jurkat whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32569 in Jurkat whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

       

    • Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)
      Anti-PI3 Kinase p110 beta antibody [Y384] (ab32569)

    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

    References (15)

    Publishing research using ab32569? Please let us know so that we can cite the reference in this datasheet.

    ab32569 has been referenced in 15 publications.

    • Hong S  et al. Ginsenoside Rg3 enhances the anticancer effect of 5-FU in colon cancer cells via the PI3K/AKT pathway. Oncol Rep 44:1333-1342 (2020). PubMed: 32945504
    • Romeo V  et al. Effects of PI3K Inhibition on Afucosylated Antibody-Driven Fc?RIIIa Events and Phospho-S6 Activity in NK Cells. J Immunol 203:137-147 (2019). PubMed: 31092639
    • Li P  et al. Expression and Comparison of Cbl-b in Lung Squamous Cell Carcinoma and Adenocarcinoma. Med Sci Monit 24:623-635 (2018). PubMed: 29384143
    • Yang Y  et al. Microarray pathway analysis indicated that mitogen-activated protein kinase/extracellular signal-regulated kinase and insulin growth factor 1 signaling pathways were inhibited by small interfering RNA against AT-rich interactive domain 1A in endometrial cancer. Oncol Lett 15:1829-1838 (2018). PubMed: 29399196
    • Si F  et al. MicroRNA-138 suppresses cell proliferation in laryngeal squamous cell carcinoma via inhibiting EZH2 and PI3K/AKT signaling. Exp Ther Med 14:1967-1974 (2017). PubMed: 28962111
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-PI3 Kinase p110 beta antibody [Y384]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (cancer cells)
    Loading amount
    15 µg
    Specification
    cancer cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 07 2007

    Question

    Hello,

    We have a quality complaint for item ab55593. Please check attached file for detail.

    We request money back.

    Thanks,


    Item number: ab55593

    Order number: 1049169



    General Information

    Antibody storage conditions？





    -20









    Description of the problem (high background, wrong band size, more bands, no band etc.)





    no band









    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)





    Human/Cell extract









    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)





    RIPA buffer containing protease and phosphatase inhibitor cocktails (Roche Diagnostics) and also heating sample







    Amount of protein loaded





    60, 80μg of total protein extracts









    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)





    10% SDS-PAGE gels









    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)





    nitrocellulose membranes (Milliopore) 90 min

    The membranes were blocked with 5% milk powder, washed with TBST







    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)





    Primeary antibody purchased from Abcom named ab55593

    Primary antibody is diluted with 4% BSA. And the membranes were incubated overnight at 4℃. The membranes were washed with TBST three times.







    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)





    peroxidase-conjugated anti-mouse IgG (Sigma) as a secondary antibody

    Incubation for 2 hours was at room temperature.

    Also washed three times by TBST.







    Detection method (ECL, ECLPlus etc.)





    Western Blotting luminal reagent (Santa cruz)







    Positive and negative controls used (please specify)





    Gapdh as positive control









    Instruction

    Repeat times: 3


    Our western blotting experiment is based on the instruction provided by your company, but we got no band. At the same time, we used the protein of 293T cell (published by some paper) to repeat the experiment and we got the same result.

    We have prolonged the time of the incubation and exposure, and also use the Concentration 1:200, but there is still no band of PI3KCB. There is no problem with our technology of western blotting, because we have got the target band of PI3KC3 by antibody from CST.

    Finally, I hope that your company can make up our loss and solve the problems as soon as possible.

    Read More
    Answer

    Thank you for your enquiry regarding ab55593 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

    I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    As the datasheet indicates this antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.

    Would you be so kind to confirm the sample types used (species, cells, tissues)?

    We have several alternative antibodies against the same target (PI3 Kinase p110 beta - ab32569, ab97322, ab1678) and I could offer you a free of charge replacement, or a credit note or a full refund as compensation.

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

    Read More

    Question

    Can you tell me if anything is known about the cross reactivity of ab32569 (PI3 Kinase p110 beta antibody) to PI3 Kinase p110 delta? Please send me the sequence of the antibody's immunogen (synthetic peptide of human PI3-K p110 beta)?

    Read More
    Answer

    Thank you for your enquiry. I am sorry but the sequence of the immunogen remains proprietary information. Rabbit monoclonal [Y384] to PI3 Kinase p110 beta (ab32569) has not been tested against the delta form of the protein. However, there appears to be very low sequence similarity between the beta peptide sequence and delta amino acid sequence. Therefore I would predict that reactivity would be highly unlikely. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

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