Overview

  • Product name
    Anti-BTK (phospho Y223) antibody
    See all BTK primary antibodies
  • Description
    Rabbit polyclonal to BTK (phospho Y223)
  • Host species
    Rabbit
  • Specificity
    ab51210 detects endogenous levels of BTK only when phosphorylated at tyrosine 223.
  • Tested applications
    Suitable for: ELISA, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthesized phosphopeptide derived from human BTK around the phosphorylation site of tyrosine 223 (A-L-YP-D-Y).

  • Positive control
    • Extracts from Hela cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab51210 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
ELISA 1/40000.
WB 1/300 - 1/1000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).

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 (ab51210)

    Lane 1 : Extracts from Hela cells treated with serum (10%, 15min) with no immunizing peptide
    Lane 2 : Extracts from Hela cells treated with serum (10%, 15min) with immunizing peptide

    Predicted band size: 76 kDa

References

ab51210 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for your reply.

We have some conjugation kits which may be of use to you in biotinylating the anti-BTK (phospho Y223) antibody (ab51210). More information on these kits can be found from the following links:

https://www.abcam.com/index.html?pageconfig=resource&rid=13148#biotin

https://www.abcam.com/index.html?pageconfig=resource&rid=13156

We have two different biotin conjugation kits, type A and type B. Both of which are available in 3x10μg, 3x100μg and 1x1mg sizes, dependent on how much antibody you are hoping to conjugate. The Type A kit is intended for assays in which a streptavidin-labeled detection reagent will be used, whilst Type B is optimised for assays in which the biotinylated protein/antibody is captured by streptavidin immobilized on a surface (ie plates, nitrocellulose, magnetic beads etc).

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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Answer

Thank you for your reply.

I would be pleased to provide you with the Abtrial testing discount codes. In order to do so, could you please confirm which distributor you will be purchasing the antibodies from?

I look forward to receiving your reply.

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Question
Answer

Thank you for contacting us and your interest in our products.

The antibodies you have chosen for your ELISA should work well together. The immunogen used to raise the mouse monoclonal ab54219 does not cover the Y223 region of BTK and they should therefore not be detecting the same epitope.

However, as we have not tested these antibodies together in a sandwich ELISA we would not be able to guarantee their compatibility. For this reason, you would be eligible to take part in our Abtrial testing scheme.

The way this scheme works is that you would purchase ab54219 and ab51210 as normal and test it in your ELISA. As long as you then share the results with us through submitting an Abreview (no matter whether the results are negative or not) you would then receive the value of ab54219 or ab51210 of a subsequent Abcam order.

More information on this offer can be found from the following link:

https://www.abcam.com/abtrail

The only constraint in this offer is that the antibody to be tested must be purchased, the Abreview submitted, and the free credit claimed within a 4 month period.

If you would like to participate in this offer please do let me know as a discount code needs to be issued prior to the purchasing of either ab54219 or ab51210.

You mentioned that you were setting up an Alphascreen. I am sorry but I did not understand how this would work. Are you just going to use one of the primary antibodies for this? If you could explain this a little more I should be able to help further.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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Question
Answer

Following up my message from yesterday, the laboratory tells me that they will not be able to provide this Anti-BTK (phospho Y223) antibody without glycerol. Please let me know if you have any questions or if we can help in any other way.

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Answer

Thank you for your enquiry. I have checked these lot numbers and found that they come from the same master stock reference 5119, they were just aliquoted at different times and should therefore behave in an identical manner. Could the small change in reactivity be due to the secondary, or the samples maybe? I'm afraid I do not have an explanation for this change. We still have 1 vial of lot 5119 in stock and if you require more I can try to contact the source and see if we can make a special order for you, please just let me know how much you would like. I hope this information helps. Please do not hesitate to contact us if you need any more information.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been experiencing problems with ab51210 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a free replacement/credit note/refund will be offered. I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. If the suggestions do not prove to be helpful, would you please be so kind to confirm the following items in order to help me better understand the cause of the problem. If you can also provide an image that would certainly help a lot. - Were the samples kept on ice or at 4oC at all times? Detecting a band of lower molecular weight would indicate that your protein has been digested. Please make sure that you had included sufficient protease inhibitor. - Did you use reducing and denaturing conditions? Was SDS included in your loading buffer and migration buffer? - Was the lysis buffer you used as strong as RIPA buffer? Please try RIPA buffer as it is the strongest and will ensure that you get all the protein of interest out. I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me again. Also, please confirm your contact/shipping address and advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you. I look forward to hearing from you again in order to resolve the matter.

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

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