Takara
login | register | order now | view cart | feedback
Products
Cell Biology


Products >  Cell_Biology >  Signal_Transduction >  EIA_Kits >  Universal_Tyrosine_Kinase_Assay_Kit

Universal Tyrosine Kinase Assay Kit

Protein Tyrosine Kinase (PTK) is an enzyme critical to the signal transduction pathways that control cell proliferation, differentiation and carcinogenesis. PTKs are classified into two familiess: membrane receptor PTKs such as insulin and EGF receptor and non-receptor linked PTKs such as Src and FAK. Membrane receptor PTKs contain three domains: the extracellular, membrane intrinsic and intracellular domains. The extracellular and membrane intrinsic domains bind to ligands such as hormones and growth factors, while the intracellular domain contains the PTK active site. Receptor PTKs transduce signals directly into the cell through ligand mediated tyrosine phosphorylation. Non-receptor linked PTKs lack the membrane intrinsic domain and are divided into three types based upon their cellular location (i.e., membrane, cytoplasmic or nuclear). Non-receptor linked PTKs participate in various functions, including signal transduction and cell-cell contact responses.

The Universal Tyrosine Kinase Assay Kit is an in vitro enzyme immunoassay kit intended for the quantitative determination of tyrosine kinase activity in either adherent or suspension cell culture extracts. The kit can also be used to measure PTK activity levels and their regulation as well as for the in vitro screening of PTK inhibitors. The kit includes a 96-well microtiter plate coated with PTK substrate, a phosphotyrosine (pY20)-horseradish peroxidase (HRP) antibody, and an HRP substrate (TMBZ). Serial dilutions of prepared cell extract may be added alongside an ATP solution to the PTK substrate-coated microtiter plate; this enables phosphorylation of tyrosine. The sample solution is then removed and the plate thoroughly washed and blocked. Following a blocking step, pY20-HRP is added, incubated, and then replaced by the TMBZ solution. Ensuing color development enables sample PTK activity to be determined by comparing sample absorbance at 450 nm with the absorbance obtained for the standard (via a plotted standard curve).

At-A-Glance Documents Images & Data Resources

Features

  • Sandwich ELISA: Quantitative determination of tyrosine kinase activity using either adherent or suspension cell culture extracts
  • Convenience: Utilizes a PTK substrate coated onto a 96-well microtiter plate, a phosphotyrosine (PY20)-horse radish peroxidase (HRP) antibody, and HRP substrate (TMBZ)
  • Sensitivity: Detects as little as 2.16 x 10-5 U/µL sample

Applications

  • Quantitative measurement of tyrosine kinase activity
  • Analysis of PTK activity regulation
  • In vitro screening of PTK inhibitors

Components

PTK substrate-immobilized microplate1 plate (8 wells x 12 strips)
Kinase reacting solution1 vial (1 x 11 mL)
40 mM ATP-2Na (lyophilized)2 vials (for 2 x 0.55 mL)
Extraction buffer1 vial (1 x 11 mL)
PTK control (lyophilized)1 vial (for 1 x 0.5 mL)
Anti-phosphotyrosine (pY20)-HRP (lyophilized)1 vial (for 1 x 5.5 mL)
Blocking solution1 vial (1 x 11 mL)
HRP substrate solution (TMBZ)1 vial (1 x 12 mL)

Storage

2–8°C

Performance Characteristics

  • Assay range: 2.16-135 x 10-5 U/µL
  • Assay sensitivity: 2.16 x 10-5 U/µL sample (= 32 fmol/well) (as Src kinase)
  • Assay duration: 1.5-3 hours
  • Specificity: gamma-phosphate residue transfer from ATP to substrate. No reactivity with phosphorylated serine or threonine.
  • Test specimen type: Suspension or adherent cell extracts.
  • Sample volume per well: 40 µL

References

1. Olgen, S. et al. (2003) J. Enz. Inh. Med. Chem. 18(6):485-490.



 
 
Products
Cat. # Product Package Size Price # of Units Select
MK410 Universal Tyrosine Kinase Assay Kit 96 Assays $282.00
 

 



Clontech is a Takara Bio Company © 2014 Privacy Policy | Terms & Conditions