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Frequently Asked Questions

View the REVERT Total Protein Stain Instructions

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Should I stain my Western blot membrane with REVERT™ Total Protein Stain before or after I dry the membrane?

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.

Can I reuse REVERT Total Protein Stain?

REVERT is intended for a single use and reusing the stain on additional membranes is not recommended. Reusing the stain will affect normalization accuracy.

Does REVERT interfere with antibody binding or the detection of my Western blot target?

REVERT is compatible with all subsequent Western blot immunodetection procedures and does not interfere with antibody binding.

How sensitive is REVERT?

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.

After staining my membrane with REVERT, I am not seeing any band development.

Here are some possible causes:

  1. Transfer was compromised. Check to see if your molecular weight ladders transferred.
  2. Membrane was not rinsed with water prior to staining.
  3. Methanol was not added to the Stain Solution.

I stained my membrane with REVERT and am getting very high background. Do you have any tips?

Prepare the Stain Solution by adding 30 mL of methanol prior to use.

How should I store REVERT Total Protein Stain and what is the shelf life?

We recommend storing REVERT Total Protein Stain at room temperature where it is stable for 6 months from the date of receipt.

Can I strip and reprobe Western blots stained with REVERT?

Yes, you can strip and reprobe the membrane for subsequent detection of another target in the 800 channel.

Can I use REVERT on a blot that was stripped?

The signal will be weak, so we do not recommend using the REVERT signal to normalize the reprobed target.

Can I stain my blot with REVERT after blocking the membrane?

No, REVERT will stain blocking buffer proteins that bind to the membrane. This signal will significantly interfere in accurately detecting the protein in the sample.

Can REVERT Total Protein Stain be detected by other imagers?

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.

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