Bio-rad Clarity™ Western ECL Substrate User Manual

ImagingCatalog # Description 170-5060 Clarity Western ECL Substrate, 200 ml 170-5061 Clarity Western ECL Substrate, 500 mlLife Science Group

6 Technical Support: 1-800-424-6723 • www.bio-rad.com4. Incubate the membrane in diluted primary antibody solution for 1 hr with continuous agitati

Technical Support: 1-800-424-6723 • www.bio-rad.com 74. Remove the membrane from the substrate solution and drain off excess.5. Place membrane i

8 Technical Support: 1-800-424-6723 • www.bio-rad.comTroubleshootingProblem Cause SolutionHigh backgroundBlocking was incomplete• Increase the conc

Technical Support: 1-800-424-6723 • www.bio-rad.com 9Troubleshooting, continuedProblem Cause SolutionNo reaction or weak signalProteins may be was

10 Technical Support: 1-800-424-6723 • www.bio-rad.comTroubleshooting, continuedProblem Cause SolutionPoor antibody binding to the antigen• Deterge

Technical Support: 1-800-424-6723 • www.bio-rad.com 11Ordering InformationCatalog Product170-5060 Clarity™ Western ECL Substrate, 200 ml size cont

Life Science Group 0712 Sig 121110026389 Rev A US/EGBio-Rad Laboratories, Inc. Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9

Table of ContentsIntroduction 1Quick Start Protocol

Technical Support: 1-800-424-6723 • www.bio-rad.com 1IntroductionThe Bio-Rad Clarity™ western ECL substrate is compatible with any HRP-conjugate s

2 Technical Support: 1-800-424-6723 • www.bio-rad.comBackground on ChemiluminescenceChemiluminescence is a chemical reaction that produces light and

Technical Support: 1-800-424-6723 • www.bio-rad.com 3Fig. 1. Specific enzymatic detection of membrane-bound antigens.

4 Technical Support: 1-800-424-6723 • www.bio-rad.comFig. 2 Chemiluminescence detection by HRP. The secondary antibodyis linked to an enzyme, which

Technical Support: 1-800-424-6723 • www.bio-rad.com 5Detailed Assay ProcedureExample Western ProtocolMaterials• PVDF, LF PVDF, or nitrocellulose