Description
General description
Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
Tyrosine phosphorylation represents a major mechanism for the activation of signal transduction pathways in response to growth factors, hormones and cytokines. There are approximately 100 tyrosine kinases that are both receptor tyrsosine kinase receptors for various growth factors and cytokines. The extent and duration of tyrosine phosphorylation are tightly regulated by the 107 protein tyrosine phosphatases. Tyrosine phosphorylation occurs on residues on either other kinases (Ser/Thr or Tyr) or on substrates. In the case of kinases, this phosphorylation often causes the enzyme to change activation states and/or it creates binding sites for SH2 domain. Mutation or misregulation of protein tyrosine kinases is a cause or consequence of many disease states. It has been estimated that 20% of oncogenes so far described are tyrosine kinases. With such clear links between aberrant protein tyrosine kinase activity and disease, it is no surprise that much of the early effort in kinase drug discovery was focused on these enzymes.
Immunogen
Phosphotyrosine conjugated to carrier protein.
Application
Anti-Phosphotyrosine Antibody, clone PY20 is an antibody against Phosphotyrosine for use in ELISA, IP & WB.
Immunoprecipitation:
Recommended by an independent laboratory to immunoprecipitate tyrosine phosphorylated proteins.
ELISA:
Recommended by an independent laboratory.
Biochem/physiol Actions
Reactivity with other species has not been confirmed.
Physical form
0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH 7.4. Liquid at 2-8°C.
Format: Purified
Preparation Note
Stable for 1 year at 2-8°C from date of recipt.
NOTE: DO NOT FREEZE.
For maximum recovery of the product, centrifuge the original vial prior to removing the cap. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8°C. Save the supernatant for application.
Analysis Note
Control
Positive Antigen Control: Catalog #12-302, EGF-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Routinely evaluated by western blot with RIPA lysates EGF-treated human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected tyrosine-phosphorylated proteins in RIPA lysates EGFtreated human A431 carcinoma cells.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Replaces: MAB3080
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
