At 09:10 AM 8/17/99 -0500, Joo Yun Hong wrote:
>hello!!
>>i'm using hydridoma supernatant to screen monoclonal antibody about
>protein on a
>membrane. since the amount of supernatant i can use is limited to a
>small volume, i
>used this apparatus named 'mini-protean2' from the bio-rad. but it has
>been giving
>me a problem of leakage between the wells. so i was wondering if anyone
>could tell
>me about similar kind of apparatus using a membrane to screen multiple
>samples of
>hydridoma supernatant or have any idea of eliminating the leakage
>problem.
>> thank
>you
>>joo jhong1 at uic.edu>
I am not too impressed with the set up of the mini protean 2. However,
leakage between the wells is not something that I have ever encountered in a
protein gel where the lanes are 'preformed' in the stack. I am not sure why
you should experience this. The only reason I can come up with is that if
the stacking gel is set too much in advance it is drying up. If the stacking
gel dries up, it pulls away from the glass plates, causing the sample to
'leak' out horizontally (it has to be poured several hours in advance for
this to happen!). The stacking layer should not be poured unless you are
ready with your samples, since it only takes about 30 min to polymerize.
BTW, if you are just screening monoclonals with hyb. sup., why not use a dot
blot?
Dr. Hiranya Sankar Roychowdhury
GENE LAB/ EPPWS
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu
