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Protein Detection and Quantitation

Introduction

Protein gel stain and Western Blot are important tool in proteomics (Figure 1). Applied BioProbes 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. In addition, Applied BioProbes also offers a phosphoprotein selective stain: Phos-Tag™ Phosphoprotein Gel Stain. Applied BioProbes’ ECL Western Blotting Detection Kit provides highly sensitive detection on immunoblots. Besides, Applied BioProbes 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 Phos-Tag™ Phosphoprotein Gel Stain
Sensitivity 20 ng 0.5 ng 1 ng
Ease of use Ready to use Supplied as stock solution Ready to use
Time required 2 hr (standard)
20 min (microwave)
90 min (standard)
30 min (microwave)
4 hr (standard)
2 hr (microwave)
Detection method colorimetric fluorescent fluorescent
MS spec. compatible Yes Yes Yes

 

Coomassie  

Figure 2. Gel stain results with Coomassie® Blue Fast Stain (left), eLuminol™ Protein Gel Stain (middle), and Phos-Tag™ Phosphoprotein Gel Stain, 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).

 

To order

Buy Catalog # Product Name Description Size List Price
P001 Coomassie® Blue Fast Stain Colorimetric protein gel stain 1 L $70

P003A
P003B
eLuminol™ Protein Gel Stain (1,000X) Fluorescent protein gel stain 500 µL
1 mL
$70
$120
P005A Phos-Tag™ Phosphoprotein Gel Stain Phosphoprotein gel stain 500 mL $180
P005B Phos-Tag™ Phosphoprotein Destain Solution Destain solution 500 mL $50
P007 ECL Western Blotting Detection Kit Western blotting reagent 100 mL $80
P010 Bradford Protein Assay Kit Protein quantitation 500 mL $75
P011 BCA Protein Assay Kit Protein quantitation 500 mL $95
P013 SDS-PAGE Sample Loading Buffer (5X) Loading buffer 5 mL $20