Product Uses Include
- Rac signaling pathway studies
- Rac activation assays with primary cells
- Studies of Rac activators and inactivators
- Rac activation assays with limited material
- High throughput screens for Rac activation
Introduction
The G-LISA series of Small G-Protein Activation Assays are ELISA based assays with which you can measure the GTP form of small G-proteins from lysates of cells or tissues and all in less than 3 h. The Rac1,2,3 G-LISA™ Activation Assay measures the entire level of GTP-loaded Rac1,2 and 3 protein in cell lysates, this is in contrast to Cat.# BK126 which measures only Rac1 activation levels. The level of activation is measured with absorbance at 490nm. For a kit to measure RhoA activation please check Cat.# BK124.
The Rac G-LISA Activation Assay is very sensitive and has excellent accuracy for duplicate samples.
Kit contents
The kit contains sufficient reagents to perform 96 Rac activation assays. Since the Rac-GTP affinity wells are supplied as strips and the strips can be broken into smaller pieces, each kit can be used for anywhere from one to multiple assays. The following components are included in the kit:
- 96 Rac-GTP affinity wells (divisible into 12 strips of 8 wells each)
- Lysis buffer
- Binding buffer
- Antigen presenting buffer
- Wash buffer
- Antibody dilution buffer
- Anti-Rac antibody
- HRP-labeled secondary antibody
- Positive control Rac protein
- Protease inhibitor cocktail (Cat. # PIC02)
- Absorbance detection reagents
- Precision Red™ Advanced protein assay reagent (Cat. # ADV02)
- Manual with detailed protocols and extensive troubleshooting guide
Equipment needed
- 96-well plate spectrophotometer capable of reading 490 nm wavelength
- Multichannel or multidispensing pipettor
- Orbital microplate shaker capable of at least 200 rpm shaking (400 rpm is optimal)
