Western Blot Storage Protocol

It may be desirable to store the western blot membrane for future analysis or reprobing. An advantage of polyvinylidene difluoride (PVDF) over nitrocellulose (NC) membranes is the stability of PVDF, which is a chemically resistant polymer that has excellent long-term stability. Thus, it is possible to store a PVDF membrane and preserve sample proteins for extended periods of time.

Blot Storage Protocol

• Protocol A
1. Sandwich the dry PVDF membrane between two clean sheets of filter paper.
2. Place the sandwich between two sheets of card stock or thin cardboard.
3. Use paperclips to clip the stack together on the edges.
4. Place the stack in a plastic bag and seal the plastic bag closed.
5. Store the PVDF membrane at 4 ˚C for up to 2 weeks, -20 ˚C for up to 2 months, or -70 ˚C for longer storage.

Note: Thaw frozen blots to RT before removal from the plastic bag because frozen blots are prone to breakage.

• Protocol B
1. PVDF membrane could also be stored wet at 4 ˚C, but sodium azide should be added to prevent bacterial growth. As sodium azide inhibits HRP activity, it should be thoroughly washed out of a blot prior to use.

More WB Resource

Western Blot Protocols & Troubleshooting & Guide

Western Blot Protocol

Western Blot Troubleshooting

Sample Preparation in Western Blot Assay

Protein Transfer from Gel to Membrane in Western Blot Assay

Running an SDS-PAGE Gel in Western Blot Assay

Western Blot Illustrated Assay

Stripping and Reprobing Protocol