PageBlue™ Protein Staining Solution - FAQs

This is most likely due to insufficient destaining time. We recommend destaining the membrane in 30% acetonitrile/20% ethanol solution for an additional 5 mins.

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

This is most likely due to a problem with the western transfer. Please confirm that the transfer buffer and transfer conditions are correct.

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

Here are possible causes and solutions:

- No protein was present in sample. Load a known amount of purified protein as a control.
- Insufficient amount of protein in sample. Load more total protein in gel.
- SDS not completely removed from gel. Wash gel more extensively before staining.

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

This is most likely due to SDS interference. We recommend increasing the number of washes and/or wash volumes before staining. Destaining the gel for 5 minutes with 25% isopropanol/10% acetic acid solution or 12% trichloroacetic acid will also help.

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

Small proteins (less than 10 kDa) are susceptible to leaching from the gel during the staining procedure and require fixation with glutaraldehyde before staining the gel with PageBlue Protein Staining Solution. Other common protein fixatives (e.g., acetic acid, isopropanol, ethanol, TCA, etc.) are not suitable for this purpose, as proteins will be washed out of the gel during the staining procedure. Please see procedure for fixation, mentioned on Page 3 of the manual.

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

The first wash step in the staining procedure is designed to remove sodium dodecyl sulfate (SDS) from the gel, because SDS interferes with staining. Native gels do not contain SDS and, therefore, this step can be omitted from native PAGE applications.

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

Fixing gel proteins with 25% isopropanol/10% acetic acid solution or 12% trichloroacetic acid (TCA) for 15 minutes can increase staining sensitivity.

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

It can detect proteins with a dynamic range of 5-500 ng, which is approximately 10 times more sensitive than traditional Coomassie R-250-based dyes.

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

The PageBlue Protein Staining Solution can be reused up to three times without noticeable loss in detection sensitivity.

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

PageBlue Protein Staining Solution contains Coomassie G-250 for protein staining on polyacrylamide gels and PVDF membranes. The rest of the composition is proprietary.

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

We recommend storing the PageBlue Protein Staining Solution 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.