Staining of proteins on SDS polyacrylamide gels and on nitrocellulose membranes by Alta, a colour used as a cosmetic.
We describe here the use of Alta, a pre-existing scarlet-red stain of cosmetic use, for staining proteins on sodium dodecyl sulfate (SDS) polyacrylamide gels, as well as for a single step staining of gels and nitrocellulose membranes during Western blot analysis. This stain, which is composed of 0.8% Crocein scarlet (brilliant crocein) and 0.2% Rhodamine B, is inexpensive, easy to use and nearly as sensitive as Coomassie Brilliant Blue (CBB) R-250. The gels can be stained in 10% Alta (2 h) and can be destained effectively only with 7% acetic acid as opposed to the conventional destainer (methanol/acetic acid/water) required for CBB-stained gels. In an alternative procedure, the proteins can be stained on the gel while electrophoresis by simply using 5% Alta in the top tank buffer and the stain can be viewed under UV-transilluminator. This procedure can also be used for Western blot analysis, as a single step procedure for staining of proteins on the gel as well as on the nitrocellulose membrane, as the stain is retained on the membrane after protein transfer. Thus, this staining procedure allows monitoring of proteins after each step in the Western blot, thereby eliminating the need to run separate gels for staining and Western blot analysis, and also the need for Ponceau Red S staining of the nitrocellulose membrane during Western blot analysis.
Pal
Jayanta K
JK
Department of Biotechnology, University of Pune, Pune 411 007, India. jkpal@unipune.ernet.in <jkpal@unipune.ernet.in>
Godbole
Dhanashri
D
Sharma
Kiran
K
D001391:Azo Compounds; D015153:Blotting, Western / Q000379:methods*; D003101:Collodion / Q000737:chemistry*; D004396:Coloring Agents; D003358:Cosmetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008567:Membranes, Artificial; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*
D001391:Azo Compounds; D004396:Coloring Agents; D003358:Cosmetics; D008567:Membranes, Artificial; D011506:Proteins; D003101:Collodion