Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)

Submit a review Q&A (4)References (5)

Overview

  • Product name

    Anti-PI 3 Kinase p110 delta antibody [EPR386]
    See all PI 3 Kinase p110 delta primary antibodies

  • Description

    Rabbit monoclonal [EPR386] to PI 3 Kinase p110 delta

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PI 3 Kinase p110 delta aa 200-300. The exact sequence is proprietary.

  • Positive control

    • WB: Jurkat, Raji, K562, RAW 264.7, Rat brain, Rat kidney and Rat spleen lysates. ICC/IF: RAW264.7 cells.

  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109006 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. Predicted molecular weight: 119 kDa.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Tissue specificity

      Expressed predominantly in leukocytes.

    • Pathway

      Phospholipid metabolism; phosphatidylinositol phosphate biosynthesis.

    • Sequence similarities

      Belongs to the PI3/PI4-kinase family.
      Contains 1 PI3K/PI4K domain.

    • Post-translational
      modifications

      Autophosphorylation on Ser-1039 results in the almost complete inactivation of the lipid kinase activity.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • 5-bisphosphate 3-kinase 110 kDa catalytic subunit delta antibody
      • 5-bisphosphate 3-kinase catalytic subunit delta isoform antibody
      • APDS antibody
      • GRB1 antibody
      • IMD14 antibody
      • p110d antibody
      • p110delta antibody
      • p110dp85a antibody
      • p85-ALPHA antibody
      • Phosphatidylinositol 3 kinase catalytic delta polypeptide antibody
      • Phosphatidylinositol 4 5 bisphosphate 3 kinase catalytic subunit delta isoform antibody
      • Phosphatidylinositol 4,5 bisphosphate 3 kinase 110 kDa catalytic subunit delta antibody
      • Phosphatidylinositol 4,5 bisphosphate 3 kinase, catalytic subunit delta antibody
      • Phosphatidylinositol 45 bisphosphate 3 kinase catalytic subunit delta isoform antibody
      • Phosphatidylinositol-4 antibody
      • Phosphoinositide 3 kinase B antibody
      • Phosphoinositide 3 kinase C antibody
      • Phosphoinositide 3 kinase catalytic delta polypeptide antibody
      • Phosphoinositide 3 kinase, catalytic, delta polypeptide variant p37delta antibody
      • PI3 kinase p110 subunit delta antibody
      • PI3-kinase subunit delta antibody
      • PI3K antibody
      • PI3K-delta antibody
      • PI3Kdelta antibody
      • Pik3cd antibody
      • PIK3R1 antibody
      • PK3CD antibody
      • PK3CD_HUMAN antibody
      • PtdIns 3 kinase p110 antibody
      • PtdIns 3 kinase subunit p110 delta antibody
      • PtdIns-3-kinase subunit delta antibody
      • PtdIns-3-kinase subunit p110-delta antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)

      Immunocytochemistry/ Immunofluorescence analysis of RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling PI 3 Kinase p110 delta with Purified ab109006 at 1:150 dilution (9.8 µg/ml). Cells were fixed in 100% Methanol and permeabilized with none. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      All lanes : Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006) at 1.5 µg/ml (purified)

      Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates
      Lane 2 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
      Lane 3 : Rat spleen lysates

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 119 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)

      Lane 1: Wild type HAP1 whole cell lysate (40 µg)
      Lane 2: PIK3CD knockout HAP1 whole cell lysate (40 µg)
      Lane 3: Jurkat whole cell lysate (40 µg)
      Lane 4: K562 whole cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109006 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified ab109006 was shown to specifically react with PIK3CD in wild-type HAP1 cells. No band was observed when PIK3CD knockout samples were used. Wild-type and PIK3CD knockout samples were subjected to SDS-PAGE. Ab109006 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

    • Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006)
      All lanes : Anti-PI 3 Kinase p110 delta antibody [EPR386] (ab109006) at 1/1000 dilution (Unpurified)

      Lane 1 : Jurkat cell lysate

      Lane 2 : Raji cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : RAW 264.7 cell lysate
      Lane 5 : Rat brain tissue lysate
      Lane 6 : Rat kidney tissue lysate
      Lane 7 : Rat spleen tissue lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 119 kDa

    References

    This product has been referenced in:

    • Cui C  et al. Znhit1 inhibits breast cancer by up-regulating PTEN to deactivate the PI3K/Akt/mTOR pathway. Life Sci 224:204-211 (2019). Read more (PubMed: 30928405) »
    • Yao Y  et al. Knockdown of long noncoding RNA Malat1 aggravates hypoxia-induced cardiomyocyte injury by targeting miR-217. Adv Clin Exp Med 28:719-728 (2019). Read more (PubMed: 30843674) »
    See all 5 Publications for this product

    Publishing research using ab109006? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Question

    Thanks for the info.
    Do you also have the concentration of Lot#: GR42675-6?

    Read More
    Answer

    That lot is also 6.8 mg/ml. Any vial labeled GR42675 will be that concentration. The number after, for instance -6 or -7, is a reference number that identifies the date the antibody was aliquotted into the vial. Please let me know if you have other questions.

    Read More

    Question

    I recently purchased the PI3kd antibody (ab109006) and I noticed that
    it is in a tissue culture supernatant format with an unknown specific
    antibody concentration. What would be the appropriate negative
    isotype control and concentration for this antibody?

    Read More
    Answer

    Thank you for contacting us.

    The IgG concentration of the lot you have, GR42675-7, is 6.8 mg/ml.

    For an isotype control, I recommend antibody ab37415:

    Click here (or use the following: https://www.abcam.com/index.html?datasheet=37415).

    The concentration of ab37415 is 5 mg/ml, so you will need to adjust the dilution factor to match the working concentration to that of ab109006.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    used ab32401 in Jurkat cells, saw no band
    received ab109006 as free of charge replacement

    for ab109006:
    Jurkat cells
    no band, but sees actin band
    Ab: 1/500
    secondary: OK
    looking for advice: use more protein, use more Ab, check if PI3K p85 can be detected

    Read More
    Answer

    Thank you for contacting us.
    I am sorry to hear that the antibodies for PI3K p110 delta do not seem to work as expected.

    As we discussed over the phone, I would like to suggest to try the following:

    1) Load moreprotein onto the gel.
    2) Use more primary antibody and incubate longer (e.g. overnight 4 degree - if you are not doing this already).
    3) If possible - check if PI3K p85 can be detected.

    Please let me know if these tips were of help. If not, please let me know as well.

    I wish you good luck and look forward to hear back from you.
    In the meantime, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    ab32401 is not detecting any bands via western blotting; secondary ab is ok; b-actin blot works well on same samples, used 1:500 with primary as suggested;

    Read More
    Answer

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab109006. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

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