Basic proteins and basic membranes adjusting blotting and staining conditions to immobilon CD

Elisabetta Gianazza, Paola Coari, Maria Rosa Lovati, Cristina Manzoni, Elena Ghibaudi, Mario Salmona

Research output: Contribution to journalArticle

2 Citations (Scopus)

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 languageEnglish
Pages (from-to)351-359
Number of pages9
JournalJournal of Chromatography A
Volume698
Issue number1-2
DOIs
Publication statusPublished - Apr 28 1995

Fingerprint

Membrane Proteins
Staining and Labeling
Membranes
Coloring Agents
Protein Sequence Analysis
Proteins
Biuret
Tolonium Chloride
Chelating Agents
Iodine
Copper
Buffers
polyvinylidene fluoride
Vapors
Degradation

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 journalArticle

Gianazza, Elisabetta ; Coari, Paola ; Lovati, Maria Rosa ; Manzoni, Cristina ; Ghibaudi, Elena ; Salmona, Mario. / Basic proteins and basic membranes adjusting blotting and staining conditions to immobilon CD. In: Journal of Chromatography A. 1995 ; Vol. 698, No. 1-2. pp. 351-359.
@article{db821a048bcb484f9862bec2b024088e,
title = "Basic proteins and basic membranes adjusting blotting and staining conditions to immobilon CD",
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.",
author = "Elisabetta Gianazza and Paola Coari and Lovati, {Maria Rosa} and Cristina Manzoni and Elena Ghibaudi and Mario Salmona",
year = "1995",
month = "4",
day = "28",
doi = "10.1016/0021-9673(94)00761-W",
language = "English",
volume = "698",
pages = "351--359",
journal = "Journal of Chromatography A",
issn = "0021-9673",
publisher = "Elsevier",
number = "1-2",

}

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 -