Validated using a knockout cell line
Recombinant
RabMAb

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

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, Flow Cyt, IPmore details
    Unsuitable for: 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
    • Jurkat whole cell lysate (ab7899).
  • General notes

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

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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 1/500. Detects a band of approximately 110 kDa (predicted molecular weight: 123 kDa).
Flow Cyt 1/40.

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

IP 1/50.
  • Application notes
    Is unsuitable for 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

    • 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.

    • 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?

    References

    This product has been referenced in:
    • Li P  et al. Expression and Comparison of Cbl-b in Lung Squamous Cell Carcinoma and Adenocarcinoma. Med Sci Monit 24:623-635 (2018). Read more (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). Read more (PubMed: 29399196) »
    See all 12 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    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
    Abcam guarantees this product to work in the species/application used in this Abreview.
    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%

    Abcam user community

    Verified customer

    Submitted Jun 07 2007

    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

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

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