Protein blotting to PVDF membranes

Andre Hamel hamel at ccu.umanitoba.ca
Mon Aug 23 11:44:52 EST 1993


In article <1993Aug23.141104.24690 at crc.ac.uk> dmartin at crc.ac.uk (David Martin x3175) writes:
>
>We routinely blot to PVDF but use 10mM CAPS pH 11 10% MeOH as the transfer
>buffer in a mini protean cell at a current of 100 mA. It takes ca. 60-90
>mins.
>
>to stain, 30 secs in coomassie (1g/l 40% MeOH 10% HOAc) then destain in
>40% MeOH 10% HOAc. repeat to enhance sensitivity (maybe use a 15 or 20 sec
>stain)
>
>we use ABI ProBlott.. and use the protocol in the instructions. Our peptide
>chemist reckons if you can see the band, it'll sequence (unless it is 
>blocked)
>
>good luck
>
>...david Martin
>
>
>dmartin at uk.ac.crc


Doesn't exposing the protein to acetic acid lead to potential acylation of
amino end? ... thus "blocking" theat end from sequencing?

                            ********************
Andre Hamel                                 email: hamel at ccu.umanitoba.ca
Manitoba Veterinary Services                lab tel.: (204) 945-7630
Infectious & Genetic Disease                     FAX: (204) 945-8062     
Mol.Biol.Lab., Winnipeg, Manitoba, CANADA
                            ********************



More information about the Methods mailing list