Need Help with detection (Western Blotting)

Protein Detection Western Blotting
1

Hi,

I was wondering what little tricks people have used in the past to align their PVDF membranes with film on exposure since my ladder doesn’t have any chemiluminesense properities and cannot be visualized on film.

Normally I would expose and then over expose to note the alignment of the PVDF membrane but I really need to be accurate. I have to identify bands which are 72 kd and 76 kd. I have used a 5-6% gel to get the adequate separation, having accuracy with film placement is just my major difficulty.

Any suggestions?

2

Laura Ripley

Simple tricks make Science what it actually is. You can mark the bands using a Pencil on Membrane after first exposure with Ponceau when you are checking the migration and then use a Sharpie to mark the bands on same position as membrane after the exposure by aligning the two.

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