After staining my PVDF membrane with PageBlue Protein Staining Solution, I am getting high background on the membrane. Can you offer some tips?
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.
After staining my PVDF membrane with PageBlue Protein Staining Solution, I am not seeing any band development. What went wrong?
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.
After staining my protein gel with PageBlue Protein Staining Solution, I am not seeing any band development. What went wrong?
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.
I stained my protein gel with Thermo Scientific PageBlue Protein Staining Solution and am getting very high background. Do you have any tips?
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.
Do you have any special recommendations for staining small proteins (less than 10 kDa) with the PageBlue Protein Staining Solution?
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.
