REVERT™ Total Protein Stain Kits for Western Blot Normalization
REVERT™ Total Protein Stain for Western Blot Normalization
REVERT™ Total Protein Stain and Wash Solution Kit
REVERT™ Wash Solution
REVERT™ Reversal Solution
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 Stain, so it is compatible with your Western blot protocol.
Total protein staining is considered the gold standard for Western blot normalization. REVERT Total Protein Stain provides linear signal over a broad range of sample concentrations and is compatible with subsequent Western blot immunodetection methods.
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. (Figure 1).
Use this stain 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 Stain provides a linear response over a wider range of cell lysate concentration than some commonly-used housekeeping proteins (Figure 2).
Biological variation won’t affect total protein normalization with REVERT. While validation of stable expression levels is highly recommended for housekeeping proteins, REVERT is a consistent internal loading control that you don’t need to validate. (Figure 3).
You can use REVERT on a membrane that was kept wet or dry following transfer. To use on a dry membrane, briefly wet the membrane in water prior to adding the Stain Solution. For PVDF membranes, rinse first in methanol, followed by water.
REVERT is intended for a single use and reusing the stain on additional membranes is not recommended. Reusing the stain will affect normalization accuracy.
REVERT is compatible with all subsequent Western blot immunodetection procedures and does not interfere with antibody binding.
REVERT is intended for staining total protein in individual lanes and has a linear detection range of 1-60 μg when used with cell lysate. However, individual proteins are easily detectable at low nanogram amounts.
Here are some possible causes:
Prepare the Stain Solution by adding 30 mL of methanol prior to use.
We recommend storing REVERT Total Protein Stain at room temperature where it is stable for 6 months from the date of receipt.
Yes, you can strip and reprobe the membrane for subsequent detection of another target in the 800 channel.
The signal will be weak, so we do not recommend using the REVERT signal to normalize the reprobed target.
No, REVERT will stain blocking buffer proteins that bind to the membrane. This signal will significantly interfere with accurate detection of the protein in the sample.
For the best results, use Odyssey imaging systems, which are designed for superior near-infrared detection. Other imagers capable of detecting signals in the 700 nm channel can also be used with the stain. Alternatively, white light imagers can detect REVERT Total Protein Stain. However, the linear and dynamic range will be reduced.