Recombinant
RabMAb

Recombinant Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217)

Rabbit recombinant monoclonal BTK (phospho Y223) antibody [EP420Y]. Validated in WB, IP, Dot and tested in Human. Cited in 9 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-BTK (phospho Y223) antibody [EP420Y]
    See all BTK primary antibodies
  • Description

    Rabbit monoclonal [EP420Y] to BTK (phospho Y223)
  • Host species

    Rabbit
  • Specificity

    BTK (phospho Y223) antibody (ab68217) detects BTK phosphorylated on tyrosine 223.

  • Tested applications

    Suitable for: WB, IP, Dot blotmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human BTK. The exact sequence is proprietary.
    Database link: Q06187
    (Peptide available as ab214589)

  • Positive control

    • WB: K562 and Ramos cell lysates treated with pervanadate. IP: Ramos cell lysates treated with pervanadate. Dot Blot: BTK (phospho Y223) phospho peptide.
  • 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 ab68217 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 1/2000 - 1/10000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).Can be blocked with BTK peptide (ab214589).

For unpurified, use 1/10000 - 1/50000.

IP 1/20.

For unpurified, use 1/40.

Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.
  • Target

    • Function

      Plays a crucial role in B-cell ontogeny. Transiently phosphorylates GTF2I on tyrosine residues in response to B-cell receptor cross-linking. Required for the formation of functional ARID3A DNA-binding complexes.
    • Involvement in disease

      Defects in BTK are the cause of X-linked agammaglobulinemia (XLA) [MIM:300755]; also known as X-linked agammaglobulinemia type 1 (AGMX1) or immunodeficiency type 1 (IMD1). XLA is a humoral immunodeficiency disease which results in developmental defects in the maturation pathway of B-cells. Affected boys have normal levels of pre-B-cells in their bone marrow but virtually no circulating mature B-lymphocytes. This results in a lack of immunoglobulins of all classes and leads to recurrent bacterial infections like otitis, conjunctivitis, dermatitis, sinusitis in the first few years of life, or even some patients present overwhelming sepsis or meningitis, resulting in death in a few hours. Treatment in most cases is by infusion of intravenous immunoglobulin.
      Defects in BTK may be the cause of X-linked hypogammaglobulinemia and isolated growth hormone deficiency (XLA-IGHD) [MIM:307200]; also known as agammaglobulinemia and isolated growth hormone deficiency or Fleisher syndrome or isolated growth hormone deficiency type 3 (IGHD3). In rare cases XLA is inherited together with isolated growth hormone deficiency (IGHD).
    • Sequence similarities

      Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
      Contains 1 Btk-type zinc finger.
      Contains 1 PH domain.
      Contains 1 protein kinase domain.
      Contains 1 SH2 domain.
      Contains 1 SH3 domain.
    • Post-translational
      modifications

      Autophosphorylated on Tyr-223 and Tyr-551. Phosphorylation of Tyr-223 may create a docking site for a SH2 containing protein.
    • Cellular localization

      Cytoplasm. Membrane. Nucleus.
    • Information by UniProt
    • Database links

    • Alternative names

      • Agammaglobulinaemia tyrosine kinase antibody
      • AGMX 1 antibody
      • AGMX1 antibody
      • AT antibody
      • ATK antibody
      • B cell progenitor kinase antibody
      • B-cell progenitor kinase antibody
      • BPK antibody
      • Bruton agammaglobulinemia tyrosine kinase antibody
      • Bruton tyrosine kinase antibody
      • Bruton’s Tyrosine Kinase antibody
      • Btk antibody
      • BTK_HUMAN antibody
      • dominant-negative kinase-deficient Brutons tyrosine kinase antibody
      • IMD 1 antibody
      • IMD1 antibody
      • MGC126261 antibody
      • MGC126262 antibody
      • OTTHUMP00000063593 antibody
      • PSCTK 1 antibody
      • PSCTK1 antibody
      • truncated Bruton agammaglobulinemia tyrosine kinase antibody
      • Tyrosine protein kinase BTK antibody
      • Tyrosine-protein kinase BTK antibody
      • tyrosine-protein kinase BTK isoform (lacking exon 14 antibody
      • XLA antibody
      see all

    Images

    • All lanes : Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217) at 1/1000 dilution

      Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate
      Lane 2 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate
      Lane 3 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate. Then the membrane was incubated with alkaline phosphatase.

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 76 kDa
      Observed band size: 76 kDa



      Blocking and dilution buffer: 5% NFDM/TBST.

    • ab68217 (purified) at 1/50 immunoprecipitating BTK (phospho Y223) in Ramos cells (Lane 1) treated with 1 mM Pervanadate for 30 min. For western blotting, a HRP-conjugated anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Dot blot analysis of BTK (phospho Y223) phospho peptide (Lane 1) and BTK phospho peptide (Lane 2) labeling BTK (phospho Y223) with ab68217 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.

    • All lanes : Anti-BTK (phospho Y223) antibody [EP420Y] (ab68217) at 1/10000 dilution (purified)

      Lane 1 : Untreated K562 cell lysate
      Lane 2 : K562 cells treated with pervanadate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size: 76 kDa
      Observed band size: 76 kDa

    References

    This product has been referenced in:

    • Herman AE  et al. Safety, Pharmacokinetics, and Pharmacodynamics in Healthy Volunteers Treated With GDC-0853, a Selective Reversible Bruton's Tyrosine Kinase Inhibitor. Clin Pharmacol Ther 103:1020-1028 (2018). Read more (PubMed: 29484638) »
    • Kim E  et al. Ibrutinib inhibits pre-BCR+ B-cell acute lymphoblastic leukemia progression by targeting BTK and BLK. Blood 129:1155-1165 (2017). WB ; Human . Read more (PubMed: 28031181) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    ELISA
    Sample
    Human Cell (Ramos cells)
    Specification
    Ramos cells
    Blocking step
    BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1.0% · Temperature: 37°C
    Type
    Sandwich (Detection)

    Biao Xi

    Verified customer

    Submitted Oct 07 2013

    Answer

    Thank you very much for your interest in ab68217.

    To our knowledge, ab68217 has not been tested in flow cytometry. Therefore, I can offer a discount off a future purchase if you buy ab68217 now, test it in flow cytometry and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and test ab68217 in flow cytometry. I will then send a discount code. This code must be issued before purchasing ab68217 so please wait for my reply before ordering.

    2. Purchase ab68217 either by phone, fax, or online (www.abcam.com).

    3. Test it in flow cytometry.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.
    Please remember that submission of the Abreview is sufficient for the discount code to become active.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab68217 turns out to be unsuitable for flow cytometry, you will still receive the discount on your next purchase after your Abreview has been submitted.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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