western blotting of acidic proteins

Sean Patterson seanpat at fmed2.uncu.edu.ar
Thu Jun 15 08:19:46 EST 2000


>Dear Netters;
>I have problem in transfering the acidic proteins (usually found in
>dentin) or phosphophoryns onto a nitrocellulose membrane. Can any one
>give me suggestions on this.
>Thankx in advance
>--
>*******************************************************************************
>*************
>
>Narayanan Karthikeyan Ph.D.,
>College of Dentistry,
>Department of Oral Biology,
>University of Illinois at Chicago
>Chicago IL 60612
>Tel: 312-413-5375
>Fax: 312-413-6694
>*******************************************************************************
>*************
>
>
>
>
>---

	An acidic protein should transfer well given it's inherent negative
charge, but might not stick well to nitrocellulose if it has no substantial
hydrophobic component.  I encountered this problem a few years ago, and
switched to a charge modified membrane. I recall that I had to block the
membrane more stringently, as it had a higher background, but otherwise
this solved my problem. Good luck.



Sean Patterson, Ph.D.
Catedra de Fisiologia, CC33
Facultad de Ciencias Medicas
Universidad Nacional de Cuyo
Mendoza, Argentina
Tel: (0261) 420-5115 ext.2684/2747
Fax: (0261) 449-4117
e-mail: seanpat at fmed2.uncu.edu.ar



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