Jurkat cell lysate stimulated with TPA was used to detect an increase in pERK expression. Revert Total Protein Stains are a reliable, effective alternative to housekeeping proteins for Western blot normalization. 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. Reliable Analysis for Quantitative Western Blotsīiological variation won’t affect total protein normalization with Revert. The relative fluorescence intensity is the average of triplicate values. C32 cell lysate was separated in 4-12% Bis-Tris gels and transferred to nitrocellulose membrane. Revert 700 remained linear from 1 to 60 μg of cell lysate. Above 20 μg of cell lysate, housekeeping proteins began to saturate and did not exhibit a linear signal response. A linear response is required for quantitation. Revert Total Protein Stains have a wider linear range than housekeeping proteins. Revert Total Protein Stains provide a linear response over a wider range of cell lysate concentration than some commonly-used housekeeping proteins (Figure 3). 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.
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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.Īccurate Normalization with Revert Total Protein Stain Revert 520 Total Protein Stain provides highly efficient protein staining on nitrocellulose or Immobilon ® FL PVDF membranes in under 10 minutes. Rapid staining with Revert 520 Total Protein Stain. 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.įigure 2. Revert 700 Total Protein Stain provides highly efficient protein staining on nitrocellulose or Immobilon ®-FL PVDF membranes in under 10 minutes. Rapid staining with Revert 700 Total Protein Stain. Revert Total Protein Stain works with your existing Western blot workflow, without needing any additional reagents or special gels. Normalization Strategies Accurate, Reliable Western Blot Normalization with Revert Stain Unlike housekeeping proteins, biological variation won’t affect total protein normalization with Revert Total Protein Staina. 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. Stain total protein in less than ten minutes on either PVDF or nitrocellulose membranes. Revert Total Protein Stains provide linear signal over a broad range of sample concentrations and are compatible with subsequent Western blot immunodetection methods. Total protein staining is considered the gold standard for Western blot normalization. Use the Gold Standard for Western Blot Normalization No special reagents, equipment, or gels are required with Revert Total Protein Stains, so they are compatible with your Western blot protocol. 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.
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