SDS-PAGE problem!

Hiranya Roychowdhury hroychow at NMSU.EDU
Mon Jul 24 10:18:09 EST 1995


On 21 Jul 1995, Dima Klenchin wrote:

> >I usually leave the extreme lanes blank
> 
> Guys,  
> 
> seems like you all missed the point. The problem is described as "curved 
> and connected bands between lanes"  and "like both ends of the bands 
> migrate faster than the middle". Note plural in both cases which 
> indicate that it is not usual smiling of the bands on edges. 
> 
Note, that in my posting I used the term 'frown', and not 'smile'. 


Clueless comments are no help.  'connected bands' result from two things: 
overload and, as another netter pointed out, slow migration of the 
polypeptides. He also suggested that 'chilled buffer' helps. Although I 
have never tried running SDS-PAGE with chilled buffer, it may help since 
chilling at 4 C does not ppt the SDS in the running buffer. 



> I never had this problem and have no clue. My guesses:
> 
> 1. 3 ug/ul is too much if you load > 5 ul.
> 2. What is salt in the samples (you decreased it less than 2-fold,
> might not be enough)?
> 3. Something wrong with running buffer (never titrate it's pH). 
> 
> -dima
> 
> 

			>>>>>>>>>>>>>>>>>>>>>>>>>>>
			  Hiranya S. Roychowdhury
   			  Plant Genetic Engineering Lab.
			  Box 3GL, NM State Univ.
			  Las Cruces, NM 88003
			  Phone: (505) 646-5785
			  hroychow at nmsu.edu
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