Bio-rad Immun-Blot® AP Colorimetric Kits Bedienungsanleitung

Immun-Blot®Assay KitInstruction ManualLIT171D 1/8/99 1:39 PM Page A

be inactivated with exposure to direct light. Warm the HRP colordevelopment buffer to RT prior to addition of the color reagentsolutions. HRP color re

3.2 Troubleshooting GuideProblem Probable cause Recommended solution1. No reaction or Color develop- Color development reagentsweak color ment solutio

Problem Probable cause Recommended solutionTransfer of protein onto themembrane was incomplete. Stainthe gel to assure transfer ofprotein. Use prestai

Section 4References1. Blake, M. S., Johnston, K. H., Russell-Jones, G. J. and Gotschlich, E. C.,Anal. Biochem., 136, 175 (1984).2. Hawkes, R., Niday,

Bio-Rad LaboratoriesLIT171 Rev DU.S. (800) 4BIORAD California (510) 741-1000 Australia 02-9914-2800 Austria (1) 877 89 01 Belgium 09-385 55 11 Canada

Table of ContentsSection 1 Preparation ... 11.1 Introduction ...

Section 1Preparation1.1 IntroductionThe Immun-Blot assay kits are enzyme immunoassay kitsoptimized for the detection of specific antigens, at picogra

Catalog Number Product DescriptionIndividual Blotting Grade Reagents (continued)170-6425 Protein G-HRP Conjugate170-6435 Premixed Tris Buffered Saline

Quantity ShelfProduct Description Provided Storage LifeComponents supplied in horseradish peroxidase kitsHRP color regent A (contains 4-chloro- 200 ml

Working solution (based on 20 assays of 5 ml each):Tris buffered saline (20 mM Tris, 500 mM NaCl, pH 7.5)(TBS) Add 100 ml of 10x TBS to a 1 L bottle a

Second antibody Prepare the second antibody solution by dissolving 33 µlconjugate solution of the antibody conjugate in 100 ml of antibody buffer, or

reactions. At 0.05%, Tween-20 will not disrupt binding of primaryantibodies to antigens or antigens to nitrocellulose, but willoptimize detection sens

Overnight incubation may be preferred, since longer incubationperiods may increase the sensitivity of detection.5. Washes - Remove the unbound first a