SDS-PAGE problem!

Hiranya Roychowdhury hroychow at NMSU.EDU
Fri Jul 21 10:13:17 EST 1995


On 20 Jul 1995, WZhao80850 wrote:

> Hello there, > I post about the same question about a month ago, until
now I haven't got > any creative suggestions and also could not solve my
problem by myself. I > am frustrating here. I problem is: > I am using
BIO-RAD mini-gel electrophoresis system. My target protein is > about
25-27 KD. I run 12% running gel and 4% stacking gel at 100 V. The >
results I've got were curved and connected bands between lanes. First I >
thought it might cause by high salt concentration in my sample, I diluted
> my sample from 5 ug/ul to 3 ug/ul.The results got better but not good >
enough. I still had curved bands and connections between lanes. It seems >
like both ends of the bands migrate faster than the middle.  > Could
anybody give me some advice? Thanks in advance! > Cheers! > Wei > > Wei
Zhao > WZhao80850 at aol.com or weiz at protein.bchs.uh.edu > Tel:
(713)669-1091(H), (713)790-4696 > Baylor College of Medicine > Houston, TX
77030 > > 

We use the same system. The 'frowning' of the extreme lanes seems to be
inherent to mini-gels (I noticed it with Hoefer's system too). However,
the problem is overcome somewhat by using thinner gels and lesser loads. 
Lesser load will also eliminate 'band-linking'. Mini-gels are limited in
applicability, especially when a low abundance protein is to be monitored. 
I usually leave the extreme lanes blank. I think the problem is with the 
construction. The extreme lanes are too close to the edge of the gel, 
where there is always a voltage drift. 
			>>>>>>>>>>>>>>>>>>>>>>>>>>>
			  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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