ProtoArray™ Kinase Substrate Identification Application Kit
from
Invitrogen
Description
The ProtoArray™ Kinase Substrate Identification Application Kit includes ProtoArray™ KSI Buffer Modules A and B. You will need to obtain a ProtoArray™ Human Protein or Yeast Proteome Microarray separately from Invitrogen before performing a microarray screening experiment.
The ProtoArray™ Human Protein Microarray is the first high-density functional protein microarray that enables you to rapidly screen your kinase against thousands of potential protein substrates in as little as two days in a simple, robust assay. Protein concentrations on the array are highly normalized compared to biological tissues enabling sensitive detection of substrates present at low concentrations.
Each ProtoArray™ Human Protein Microarray mg v1.0 contains greater than 1,800 open reading frames (ORFs) representing multiple gene families. Each ORF is expressed as a 5'-GST fusion, purified and spotted in duplicate on a 1 inch x 3 inch modified glass slide. To increase the likelihood of native structure and proper functionality, proteins were expressed in a baculovirus expression system and purified under native conditions.
Using the ProtoArray™ Human Protein Microarray for kinase substrate identification (KSI)takes four simple steps:
Prepare a kinase reaction mixture containing 33P--ATP.
Transfer the reaction to a ProtoArray™ Human Protein Microarray.
Wash and expose array to phosphor storage screen or film.
Analyze results.
Each ProtoArray™ Human Protein Microarray mg v1.0 contains greater than 1,800 unique human proteins arrayed in duplicate on a 1 inch x 3 inch modified glass microarray. • Each ProtoArray™ Kinase Substrate Identification Application Kit contains two modules: ProtoArray™ KSI Buffer Module A contains 30% BSA, 400 µl 1M DTT, Kinase Buffer (100 mM MOPS, pH 7.2, 1% NP40, 100 mM NaCl, 10 mg/ml BSA, 5mM MgCl
2, 5mM MnCl
2), 10 µl PKA Control Kinase. ProtoArray™ KSI Buffer Module B contains 14 ml 10X PBS, 20 ml 10% SDS, and 5 Hybrislip™ cover slips. • The complete kit includes two ProtoArray™ Human Protein Microarrays mg v1.0, 2 ProtoArray™ Control Protein Microarrays, and the ProtoArray™ Substrate Identification Application Kit.