Problem with fluorescein

Serguei Golovan sgolovan at uoguelph.ca
Tue Aug 29 18:40:32 EST 1995


Dear netters.
I am using fluorescein (Coul and Pappin method) for staining proteins 
transfered to PVDF membrane. 
The problem is that the staining is very poor, I have great trouble with 
localizing even 100pM of pure horse myoglobin after transfer. The 
transfer  itself seems fine and if I stain with Coumassie, I can see a 
very strong band. Also if I stain protein spotted on PVDF (dot blot), 
fluorescein works just fine,it even more sensitive than Coumassie. But 
after transfer it is totally opposite. It seems that after transfer the 
protein is somehow blocked for fluorescein.
Does anyone have any suggestions for my problem with fluorescein?
Thank you for your comments.
P.S. I did transfer with Towbins and CAPS with same results and I washed 
membrane in water and dry it very well before staining.
                                                       Sergei Golovan.
                                                       sgolovan at uoguelph.ca
 




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