Phosphotyrosine BTK ELISA Kit (ab279750)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phosphotyrosine BTK ELISA Kit
See all BTK kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phosphotyrosine BTK ELISA Kit (ab279750) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated BTK in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.
This sandwich-based ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-BTK. An anti-BTK antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated BTK present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and biotinylated anti-phosphotyrosine antibody is used to detect only tyrosine-phosphorylated protein. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phospho-BTK bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 5X Assay Diluent 1 x 15ml 600X HRP-Streptavidin Concentrate 1 vial Biotinylated anti-phosphotyrosine antibody 2 vials Pan-BTK Coated Microplate 1 unit Positive Control - THP-1 cell lysate 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
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
modificationsAutophosphorylated 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
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Alternative names
- Agammaglobulinaemia tyrosine kinase
- AGMX 1
- AGMX1
see all -
Database links
- Entrez Gene: 695 Human
- Omim: 300300 Human
- SwissProt: Q06187 Human
- Unigene: 159494 Human