Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4254

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Rapid Measurement of Protein Concentration by Western Analysis Using Colorimetric Detection by BCIP-NBT

This protocol was adapted from "Rapid Dot-Blot Western Analysis Using Nitrocellulose and Colorimetric Detection by BCIP-NBT," contributed by Joint ProteomicS Laboratory (JPSL) of the Ludwig Institute for Cancer Research and the Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia, Appendix 2, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2004.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol uses antibodies that recognize epitopes carried by the target protein to estimate the relative amounts of the protein in a series of samples. The concentration of purified protein can be measured accurately by assaying a series of standards containing known amounts of the protein of interest. When working at its best, the system is capable of detecting 100 pg of immobilized alkaline phosphatase-conjugated antibody. To detect as little as 1 . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


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