TYR Phosphorylation Assay Kit
The TYR Phosphorylation Assay Kit is designed for measuring TYR (tyrosine) phosphorylation and screening and profiling small molecules that affect it. This kit comes in a convenient 96-well format, with enough antibody specific for phosphorylated TYR residues, HRP-labeled antibody and assay buffer for 100 reactions. This kit also includes phosphorylated JAK2 (Janus kinase 2) as standard control.
- Recombinant protein of interest
- TBST Buffer (1x Tris-buffered saline, pH 8.0, containing 0.05% Tween-20)
- Luminometer or microplate reader capable of reading chemiluminescence.
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| 52140Z5 | Primary Antibody 30 | 12.5 µl | -80°C |
| 52130H | Secondary HRP-Labeled Antibody 1 | 10 µl | -80°C |
| 1x Phosphorylation Buffer | 3 x 1 ml | -20°C | |
| 79956 | Blocking Buffer | 50 ml | +4°C |
| Phosphorylated JAK2* | 50 µl | -80°C | |
| HRP Chemiluminescence Substrate A (translucent bottle) | 6 ml | +4°C | |
| HRP Chemiluminescence Substrate B (brown bottle) | 6 ml | +4°C | |
| 96-well strip plate (coated) | 1 plate | Room Temp |
* The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
TYR (tyrosine) phosphorylation is a modification of proteins where a phosphate group is added to the amino acid tyrosine by tyrosine kinases. This modification is crucial to regulate enzyme activity and signaling pathways, such as the Ras-MAPK (mitogen activated protein kinase) signaling pathway, and is needed for proteins involved in cell differentiation, cell cycle, gene regulation and transcription, angiogenesis and many other functions. Tyrosine kinases can be receptor or non-receptor, with RTK (receptor tyrosine kinases) being transmembrane proteins that can bind ligands extracellularly, while NRTK are mostly soluble intracellular proteins. Tyrosine phosphorylation can be reverted by PTPs (tyrosine phosphatases). Abnormal tyrosine phosphorylation is linked to several diseases, such as cancer and diabetes. An understanding of the TYR phosphorylation state of a protein and how to manipulate it, via the partners involved, is critical for the development of targeted therapies for tyrosine phosphorylation linked diseases.
Wang, J.Y.J. 1988. Antibodies for phosphotyrosine: Analytical and preparative tool for tyrosyl-phosphorylated proteins. Anal. Biochem.172(1): 1-7.
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