Removal of coomassie blue precipitates from polyacrylamide gels.

نویسنده

  • S A Lewis
چکیده

During staining of polyacrylamide gels, varying amounts of Coomassie blue may precipitate on the surface of the gels. This background obscures faint bands and is esthetically unappealing. The problem is worse when gels are stained for longer periods of time or when the stain has not been recently filtered. Rinsing a gel with methanol efficiently and quickly removes this background. Basically, at any time during destaining, gels are treated for 1 min in 100% methanol and returned to the destain or water. Gentle shaking is required. If the methanol rinse is done before excess stain is completely removed, it is easier to judge when destaining is complete (because of the lower background). During the treatment, gels may turn opaque, although this does not appear to harm the gel. For illustration, a 0.75-mm-thick sodium dodecyl sulfate 10% polyacrylamide gel (1) was stained with 0.25% Coomassie Brilliant Blue R®-250 (BioRad, Hercules, CA. USA) in 25% isopropanol and 10% acetic acid for 24 h and then destained in 7% acetic acid with four changes of destain over 24 h (2). The gel was cut in half. One half was kept in destain, while the other was rinsed in methanol for 1 min (Figure 1), as described above. Ethanol and acetone removed precipitated Coomassie stain at about the same rate as methanol. Isopropanol was slightly less effective. Neither 0.75-mm gels nor 1.5-mm gels (10% polyacrylamide) appeared to be harmed by 5min treatments with 100% methanol, although the stain came off the gel surface in less than one minute. Protracted incubation will remove stain from protein bands. Also, 0.75-mm 5% and 20% polyacrylamide gels appeared unharmed by the methanol treatment.

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عنوان ژورنال:
  • BioTechniques

دوره 21 5  شماره 

صفحات  -

تاریخ انتشار 1996