Abstract
Transfer efficiency from polyacrylamide gels and binding to Immobilon P and CD were tested in different buffers with 125I-labelled proteins. With a derivatized poly(vinylidene difluoride) membrane, a pH 8 medium was found to be superior to a more alkaline solution, for both acidic and basic proteins. New staining protocols were tried on Immobilon CD. Toluidine Blue and iodine vapour gave a negative and a positive stain, respectively, with a fair band-to-background contrast. Protein sequencing after both stains was not impaired by interfering peaks. Biuret solution stained the protein bands pale pink but, even after copper removal with a chelating agent, it completely prevented Edman degradation. The first two procedures compare favourably with a commercial kit for protein detection, Quick Stain, that provides comparable sensitivity but results in several spurious peaks on protein sequencing.
| Original language | English |
|---|---|
| Pages (from-to) | 351-359 |
| Number of pages | 9 |
| Journal | Journal of Chromatography A |
| Volume | 698 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - Apr 28 1995 |
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ASJC Scopus subject areas
- Biochemistry
- Analytical Chemistry
- Organic Chemistry
Cite this
Basic proteins and basic membranes adjusting blotting and staining conditions to immobilon CD. / Gianazza, Elisabetta; Coari, Paola; Lovati, Maria Rosa; Manzoni, Cristina; Ghibaudi, Elena; Salmona, Mario.
In: Journal of Chromatography A, Vol. 698, No. 1-2, 28.04.1995, p. 351-359.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Basic proteins and basic membranes adjusting blotting and staining conditions to immobilon CD
AU - Gianazza, Elisabetta
AU - Coari, Paola
AU - Lovati, Maria Rosa
AU - Manzoni, Cristina
AU - Ghibaudi, Elena
AU - Salmona, Mario
PY - 1995/4/28
Y1 - 1995/4/28
N2 - Transfer efficiency from polyacrylamide gels and binding to Immobilon P and CD were tested in different buffers with 125I-labelled proteins. With a derivatized poly(vinylidene difluoride) membrane, a pH 8 medium was found to be superior to a more alkaline solution, for both acidic and basic proteins. New staining protocols were tried on Immobilon CD. Toluidine Blue and iodine vapour gave a negative and a positive stain, respectively, with a fair band-to-background contrast. Protein sequencing after both stains was not impaired by interfering peaks. Biuret solution stained the protein bands pale pink but, even after copper removal with a chelating agent, it completely prevented Edman degradation. The first two procedures compare favourably with a commercial kit for protein detection, Quick Stain, that provides comparable sensitivity but results in several spurious peaks on protein sequencing.
AB - Transfer efficiency from polyacrylamide gels and binding to Immobilon P and CD were tested in different buffers with 125I-labelled proteins. With a derivatized poly(vinylidene difluoride) membrane, a pH 8 medium was found to be superior to a more alkaline solution, for both acidic and basic proteins. New staining protocols were tried on Immobilon CD. Toluidine Blue and iodine vapour gave a negative and a positive stain, respectively, with a fair band-to-background contrast. Protein sequencing after both stains was not impaired by interfering peaks. Biuret solution stained the protein bands pale pink but, even after copper removal with a chelating agent, it completely prevented Edman degradation. The first two procedures compare favourably with a commercial kit for protein detection, Quick Stain, that provides comparable sensitivity but results in several spurious peaks on protein sequencing.
UR - http://www.scopus.com/inward/record.url?scp=0028942886&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028942886&partnerID=8YFLogxK
U2 - 10.1016/0021-9673(94)00761-W
DO - 10.1016/0021-9673(94)00761-W
M3 - Article
AN - SCOPUS:0028942886
VL - 698
SP - 351
EP - 359
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 1-2
ER -