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Protein Detection and Quantitation

Introduction

Protein gel staining and Western Blotting are important tools in proteomics (Figure 1). GeneCopoeia offers two distinct protein gel stains: 1) Coomassie® Blue Fast Stain, a ready-to-use product that does not require methanol or acetic acid fixatives or destains; 2) eLuminol™ Protein Gel Stain, a highly-sensitive fluorescent stain designed to detect proteins in SDS polyacrylamide gels.  GeneCopoeia’ ECL Western Blotting Detection Kit provides highly sensitive detection on immunoblots. Besides, GeneCopoeia also offers two protein assay kits: 1) Bradford Protein Assay Kit; and 2) BCA Protein Assay Kit.

Figure 1. Protein gel analysis workflow

Protein Gel Stain Selection Guide

  Coomassie® Blue Fast Stain eLuminol™ Protein Gel Stain
Sensitivity 20 ng  0.5 ng
Ease of use Ready to use Supplied as stock solution
Time required 2 hr (standard)
20 min (microwave)
90 min (standard)
30 min (microwave)
Detection method colorimetric fluorescent
MS spec. compatible       Yes Yes

 

Coomassie  

Figure 2. Gel stain results with Coomassie® Blue Fast Stain (left), eLuminol™ Protein Gel Stain (middle), followed by eLuminol™ total protein stain (right). Two-fold serial dilutions (from left to right) of protein marker were separated via SDS-PAGE and then stained with above stains.

ECL Western Blotting Detection Kit

Figure 3. Detection of protein in Western Blots with ABP kit (left) and Millipore Kit (right).

                                      Bradford Protein Assay Kit  BCA Protein Assay Kit

Figure 4. BSA standard curve with Bradford Protein Assay Kit (left) and BCA Protein Assay Kit (right).