Pierce™ Reversible Protein Stain Kit for PVDF Membranes - FAQs

The procedure for the Reversible Protein Stain Kit for PVDF Membranes (Cat. No. 24585) is performed before the membrane is blocked with the blocking buffer, and thus, has no effect on the subsequent blocking.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The following are possible causes for weak signal intensity:
- Low level of protein transferred from the gel to the membrane. This is particularly true for high molecular weight proteins. Reversible Protein Stain Kits for Nitrocellulose and PVDF membranes (Cat. Nos. 24580 and 24585, respectively) may be used to stain the bands on the membrane to indicate transfer efficiency.
- Over-diluted primary and secondary antibody probes. We suggest using the recommended dilution for the antibody probes for each substrate. More sensitive substrates will require a more dilute antibody solution.
- Short substrate incubation time. For best results, incubate the membrane for 5-15 min in the SuperSignal substrate.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

The following are possible causes for weak signal intensity:
- Low level of protein transferred from the gel to the membrane. This is particularly true for high molecular weight proteins. Reversible Protein Stain Kits for Nitrocellulose and PVDF membranes (Cat. Nos. 24580 and 24585, respectively) may be used to stain the bands on the membrane to indicate transfer efficiency.
- Over-diluted primary and secondary antibody probes. We suggest using the recommended dilution for the antibody probes for each substrate. More sensitive substrates will require a more dilute antibody solution.
- Short substrate incubation time. For best results, incubate the membrane for 5-15 min in the SuperSignal substrate.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Here are possible causes and solutions:

- Membrane was allowed to dry before reversing. Keep the membrane wet.
- High protein concentration in the sample. Extend incubation in the Stain Eraser up to 5 minutes. Reduce the protein concentration in the sample.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

This is likely due to low amounts or no protein present in the sample. Please determine the protein concentration in the original sample.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes detects as low as 25 to 50 ng protein per band.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

We recommend storing the MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes at room temperature where it is stable for a year.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.