Human Phospho-Receptor Tyrosine Kinase Array Kit

Introduction

R&D Systems™ Human Phospho-Receptor Tyrosine Kinase (RTK) Array Kit (Catalog # ARY001B) is a screening tool designed to simultaneously detect the relative phosphorylation of 49 different RTKs. This kit eliminates the need for numerous immunoprecipitation (IP) and/or Western blot experiments. RTKs are captured by antibodies spotted on a nitrocellulose membrane. Levels of Phospho-RTK are then assessed using an HRP-conjugated pan phospho-tyrosine antibody followed by chemiluminescence detection.

Simultaneously detect the relative phosphorylation of these RTKs in a single sample
ALK/CD246 EphB4 MuSK
Axl EphB6 PDGF R alpha
DDR1 ErbB2 PDGF R beta
DDR2 ErbB3 c-Ret
Dtk ErbB4 ROR1
EGF R FGF R1 ROR2
EphA1 FGF R2 alpha Ryk
EphA2 FGF R3 SCF R/c-kit
EphA3 FGF R4 Tie-1
EphA4 Flt-3/Flk-2 Tie-2
EphA5 HGF R/c-MET TrkA
EphA6 IGF-I R TrkB
EphA7 Insulin R/CD220 TrkC
EphA10 M-CSF R VEGF R1/Flt-1
EphB1 Mer VEGF R2/KDR
EphB2 MSP R/Ron VEGF R3/Flt-4
EphB3    

General Assay Principle

Carefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell lysate samples are diluted and mixed with a cocktail of biotinylated detection antibodies. The sample/antibody mixture is then incubated with the array. Any protein/detection antibody complex present is bound by its cognate immobilized capture antibody on the membrane. Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of cytokine bound. Chemiluminescence is detected in the same manner as a Western blot.

Kit Contents

  • 4 Array Membranes
  • 4-Well Multi-dish
  • Array Buffers
  • Lysis Buffer
  • Wash Buffer
  • Anti-Phospho-Tyrosine-HRP Detection Antibody
  • Chemiluminescent Detection Reagents
  • Transparency Overlay Template
  • Detailed Protocol

For a complete list of the kit contents and necessary materials, please see the Materials Provided/Other Supplies Required sections of the product datasheet.