TY - JOUR
T1 - Staining protein in isoelectric focusing gels with fast green
AU - Allen, Ronald E.
AU - Masak, Kenneth C.
AU - McAllister, Pamela K.
N1 - Funding Information:
This work is Michigan Agricultural Experiment Station Article No. 9206, and was supported in part by Michigan Agricultural Experiment Station Project 1280 and 1182, a biomedical research support grant from the College of Agriculture and Natural Resources. and USPHS Grant lR23 AG01467-01.
PY - 1980/5/15
Y1 - 1980/5/15
N2 - A rapid, simple technique for staining proteins in isoelectric focusing polyacrylamide gels was demonstrated using fast green in 10% acetic acid. Fast green has the distinct advantage of not binding to ampholytes, thus staining only protein. Maximum staining was achieved within 5 min, and bands were visible after 3 to 6 h of destaining. Background stain removal, however, was not complete until 72 h after placing gels in a diffusion destainer. Gel quantitation was demonstrated with actin using fast green and Coomassie brilliant blue R-250. A standard curve prepared with fast green was linear from 0.5 to 8 μg of actin in contrast to Coomassie brilliant blue R-250 which provided linearity from 0.1 to 2.5 μg actin. Application of fast green staining to quantitation of α-actin from cultured muscle satellite cells has been demonstrated.
AB - A rapid, simple technique for staining proteins in isoelectric focusing polyacrylamide gels was demonstrated using fast green in 10% acetic acid. Fast green has the distinct advantage of not binding to ampholytes, thus staining only protein. Maximum staining was achieved within 5 min, and bands were visible after 3 to 6 h of destaining. Background stain removal, however, was not complete until 72 h after placing gels in a diffusion destainer. Gel quantitation was demonstrated with actin using fast green and Coomassie brilliant blue R-250. A standard curve prepared with fast green was linear from 0.5 to 8 μg of actin in contrast to Coomassie brilliant blue R-250 which provided linearity from 0.1 to 2.5 μg actin. Application of fast green staining to quantitation of α-actin from cultured muscle satellite cells has been demonstrated.
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U2 - 10.1016/0003-2697(80)90106-2
DO - 10.1016/0003-2697(80)90106-2
M3 - Article
C2 - 6160787
AN - SCOPUS:0019308023
SN - 0003-2697
VL - 104
SP - 494
EP - 498
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -