SDS Gel

Ctro Inmunologia Molecular monoclon%infomed at GN.APC.ORG
Fri Sep 30 23:30:49 EST 1994


   
 
   Dear Paul 
 
   
 
   A simple non-destructive method for detecting proteins in sds gel 
 
   prior to elution and further analysis is to incubate the gel in 0.25 
 
   M KCL.The band can then be cut out,the gel crushed,and the protein 
 
   eluted eith a buffer solution containing sds 0.1%;the protein is then 
 
   concentrated and sds remove by dialisis.
 
   
 
   Another method is stain the gel with Imidazole-Zinc salts like 
 
   follow( Biotechniques 565 -573):
 
   
 
   1-Rinse:place the gel in deionized water for 5 
 
   s. following sds-page.
 
   2-Equilibration:soak the gel in 50-100 ml 0.2 M imidazole in water 
 
   with gentle shaking. Recommended times:10% acrylamide concentration 5 
 
   min.; for 12.5 % acrylamide,5-10 min.;and equal for greater than 15 
 
   %,15-30 min.
 
   3-Staining:Immerse gel in 50-100 ml of 0.3 M zinc or copper salts 
 
   (sulfate,chloride,acetate ) and shake gently for one min.Discard the 
 
   staining solution when background becomes deep white or dark 
 
   blue,respectivley,with transparent proteins bands.
 
   4-Storage:store reverse -stained gel either in deionized water or in 
 
   sealed polyethylene bags.
 
   
 
   References:
 
    1-Hager,D.A.&Burgess,R.R.,Anal.Biochem.109,76-86 ( 1980 )
 
    2-Dzandu,J.K. et al. Anal.Biochem.174:157-167 (1988)
 
    3-Lee,C. et al .Anal.Biochem. 166:308-312 (1987)
 
    4-Biotechniques vol.12,No. 4 ( 1992 )
 
       
 
   
 
    Good Luck, Paul
 
    
 
    Lobvi E. Matamoros Fernandez
 
    Quality Control Unit
    Center of Molecular Immunlogy.
 
    e. mail: monoclon at infomed.sld.cu
    PO BOX: 16040, Havana 11600, CUBA
   



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