Human Phospho-Immunoreceptor Array Kit

• Human Phospho-Immunoreceptor Array Kit Ordering Information

Introduction

R&D Systems Human Phospho-Immunoreceptor Array (Catalog # ARY004B) is a rapid, sensitive, and economical tool used to simultaneously detect the relative phosphorylation levels of 59 ITAM/ITIM-associated immunoreceptors, avoiding numerous immunoprecipitations and/or Western blots.

• Sample Data
• Troubleshooting Guide

General Assay Principle

Carefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell lysate samples are diluted and incubated with the array. Any protein present in the sample is bound by its cognate immobilized capture antibody on the membrane. A pan anti-phospho-tyrosine antibody conjugated to horseradish peroxidase (HRP) is then used to detect phosphorylated tyrosines on activated receptors. Chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of protein bound. Chemiluminescence is detected in the same manner as a Western blot.

Alternative Total Immunoreceptor Profile Assay Principle

This kit can also be used to obtain a total immunoreceptor profile by using lysates prepared from cell-surface biotinylated cells and Streptavidin-HRP (Catalog # DY998) for detection. A biotinylated positive control for streptavidin-HRP detection is spotted on three corners of the array next to the phospho-tyrosine positive control. Details for this protocol can be found by clicking the link below.

• Cell-Surface Biotinylation Profile Protocol

Kit Contents

• 4 Array Membranes
• 4-Well Multi-dish
• Array Buffers
• Lysis Buffer
• Wash Buffer
• HRP-conjugated Anti-phospho-tyrosine 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.