Revert Total Protein Stain for Accurate Western Normalization

Revert™ Total Protein Stains for Western Blot Normalization

Products

Revert™ 700 Total Protein Stain for Western Blot Normalization

Revert™ 700 Total Protein Stain Kits for Western Blot Normalization

Revert™ 700 Total Protein Stain and Wash Solution Kit

Revert™ 520 Total Protein Stain for Western Blot Normalization

Revert™ 520 Total Protein Stain Kits for Western Blot Normalization

Revert™ 520 Total Protein Stain and Wash Solution Kit

Make Western blot normalization more accurate and reliable with Revert Total Protein Stain, a membrane-based (post-transfer) normalization strategy that stains all protein in your sample. No special reagents, equipment, or gels are required with Revert Total Protein Stains, so they are compatible with your Western blot protocol.

Use the Gold Standard for Western Blot Normalization

Total protein staining is considered the gold standard for Western blot normalization. Revert Total Protein Stains provide linear signal over a broad range of sample concentrations and are compatible with subsequent Western blot immunodetection methods.

  • Quick and Compatible. Stain total protein in less than ten minutes on either PVDF or nitrocellulose membranes.
  • Accurate Normalization. With a wide linear range of 1 - 60 μg, it’s easy to detect Revert stains and your targets in the same linear range for accurate normalization.
  • Reliable Analysis. Unlike housekeeping proteins, biological variation won’t affect total protein normalization with Revert Total Protein Staina.

Revert 700 Total Protein Stain Workflow

Revert 520 Total Protein Stain Workflow

Normalization Strategies

Accurate, Reliable Western Blot Normalization with Revert Stain

Revert Total Protein Stain works with your existing Western blot workflow, without needing any additional reagents or special gels. Staining takes less than ten minutes. (Figures 1 and 2).

Figure 1. Rapid staining with Revert 700 Total Protein Stain. Revert 700 Total Protein Stain provides highly efficient protein staining on nitrocellulose or Immobilon®-FL PVDF membranes in under 10 minutes. 10 μg of Jurkat cell lysate was loaded in each lane and separated in 4-12% NuPAGE Bis-Tris gels, transferred to nitrocellulose or PVDF membrane, and then imaged with the Odyssey® CLx Imaging System in the 700 nm channel.
revert 520, PVDF and nitrocellulose SD851-112
Figure 2. Rapid staining with Revert 520 Total Protein Stain. Revert 520 Total Protein Stain provides highly efficient protein staining on nitrocellulose or Immobilon® FL PVDF membranes in under 10 minutes. A dilution series using 20 µg to 313 ng of mouse heart tissue lysate was separated in 4-12% NuPAGE Bis-Tris gels, transferred to nitrocellulose or PVDF membranes, and then imaged using the 520 nm channel of an Odyssey M Imager.

Accurate Normalization with Revert Total Protein Stain

Use Revert stains to normalize accurately, by correcting for lane-to-lane variation in loading and transfer of target protein in your quantitative Western blots. Revert Total Protein Stains provide a linear response over a wider range of cell lysate concentration than some commonly-used housekeeping proteins (Figure 3).

Figure 3. Revert Total Protein Stains have a wider linear range than housekeeping proteins. A linear response is required for quantitation. Above 20 μg of cell lysate, housekeeping proteins began to saturate and did not exhibit a linear signal response. Revert 700 remained linear from 1 to 60 μg of cell lysate. C32 cell lysate was separated in 4-12% Bis-Tris gels and transferred to nitrocellulose membrane. The relative fluorescence intensity is the average of triplicate values.

Reliable Analysis for Quantitative Western Blots

Biological variation won’t affect total protein normalization with Revert. While validation of stable expression levels is highly recommended for housekeeping proteins, Revert Total Protein Stains are a consistent internal loading control that you don’t need to validate. (Figure 4).

Figure 4. Revert Total Protein Stains are a reliable, effective alternative to housekeeping proteins for Western blot normalization. Jurkat cell lysate stimulated with TPA was used to detect an increase in pERK expression. (A) Target signals were normalized to Revert 700 Total Protein Stain, which was detected in the 700 nm channel of the Odyssey CLx Imaging System. (B) Housekeeping protein normalization with tubulin, detected in the 800 nm channel. (C) Normalized results for Revert stain and tubulin were similar for pERK detection.