Proteomic capacity of recent fluorescent dyes for protein
staining

J 2004

Proteomic capacity of recent fluorescent dyes for protein staining

CHEVALIER, Francois, Valérie ROFIDAL, Pavlína VÁŇOVÁ, Alexis BERGOIN, Michel ROSSIGNOL et. al.

Basic information

Original name

Proteomic capacity of recent fluorescent dyes for protein staining

Authors

CHEVALIER, Francois, Valérie ROFIDAL, Pavlína VÁŇOVÁ, Alexis BERGOIN and Michel ROSSIGNOL

Edition

Phytochemistry, Elsevier Science, 2004, 0031-9422

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

France

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.101

Organization unit

Faculty of Education

UT WoS

000223440400004

Keywords in English

Protein staining; Fluorescent dyes; Two-dimensional electrophoresis; Proteomics

Abstract

V originále

Staining of two-dimensional gel constitutes a crucial step in comparative proteome analysis with respect to both the number of proteins analysed, the accuracy of spot quantification and reproducibility. In this work, we compared the efficiency of recent fluorophores to stain Arabidopsis total protein extract: Sypro Ruby (SR), Deep Purple (DP) and 5-hexadecanoylamino-fluorescein (C16-F). In addition, classical visible dyes, colloidal Coomassie blue (CCB) and silver nitrate (SN), were also included. High quality images were obtained for the three fluorescent dyes, DP giving the cleaner background, whereas spikes were observed with SR and a rough background with C16-F. On the other hand, saturation occurred for abundant spots with SR and DP. For a same protein load the number of detected spots ranged between 250 for CCB and 800 for SR in the sequence SR>DP>SN>C16-F>CCB. These differences were shown to rely mainly on the sensitivity between dyes leading to the detection of additional spots belonging to classes of lower abundance. Analysis of the distribution of variation coefficients for spots from replicates showed differences in the staining reproducibility between dyes that ranged in the order SR>C16-F>DP>SN>CCB. The implications of these results for the selection of a convenient stain are discussed according to specific objectives as well as practical aspects.

Citovat

CHEVALIER, Francois, Valérie ROFIDAL, Pavlína VÁŇOVÁ, Alexis BERGOIN and Michel ROSSIGNOL. Proteomic capacity of recent fluorescent dyes for protein staining. Phytochemistry. Elsevier Science, 2004, vol. 65, No 11, p. 1499-1506. ISSN 0031-9422.

@article{562182,
   author = {Chevalier, Francois and Rofidal, Valérie and Váňová, Pavlína and Bergoin, Alexis and Rossignol, Michel},
   article_number = {11},
   keywords = {Protein staining; Fluorescent dyes; Two-dimensional electrophoresis; Proteomics},
   language = {eng},
   issn = {0031-9422},
   journal = {Phytochemistry},
   title = {Proteomic capacity of recent fluorescent dyes for protein staining},
   url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15276447},
   volume = {65},
   year = {2004}
}

TY  - JOUR
ID  - 562182
AU  - Chevalier, Francois - Rofidal, Valérie - Váňová, Pavlína - Bergoin, Alexis - Rossignol, Michel
PY  - 2004
TI  - Proteomic capacity of recent fluorescent dyes for protein staining
JF  - Phytochemistry
VL  - 65
IS  - 11
SP  - 1499-1506
EP  - 1499-1506
PB  - Elsevier Science
SN  - 00319422
KW  - Protein staining
KW  - Fluorescent dyes
KW  - Two-dimensional electrophoresis
KW  - Proteomics
UR  - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15276447
N2  - Staining of two-dimensional gel constitutes a crucial step in comparative proteome analysis with respect to both the number of proteins analysed, the accuracy of spot quantification and reproducibility. In this work, we compared the efficiency of recent fluorophores to stain Arabidopsis total protein extract: Sypro Ruby (SR), Deep Purple (DP) and 5-hexadecanoylamino-fluorescein (C16-F). In addition, classical visible dyes, colloidal Coomassie blue (CCB) and silver nitrate (SN), were also included. High quality images were obtained for the three fluorescent dyes, DP giving the cleaner background, whereas spikes were observed with SR and a rough background with C16-F. On the other hand, saturation occurred for abundant spots with SR and DP. For a same protein load the number of detected spots ranged between 250 for CCB and 800 for SR in the sequence SR>DP>SN>C16-F>CCB. These differences were shown to rely mainly on the sensitivity between dyes leading to the detection of additional spots belonging to classes of lower abundance. Analysis of the distribution of variation coefficients for spots from replicates showed differences in the staining reproducibility between dyes that ranged in the order SR>C16-F>DP>SN>CCB. The implications of these results for the selection of a convenient stain are discussed according to specific objectives as well as practical aspects.
ER  -

CHEVALIER, Francois, Valérie ROFIDAL, Pavlína VÁŇOVÁ, Alexis BERGOIN and Michel ROSSIGNOL. Proteomic capacity of recent fluorescent dyes for protein staining. \textit{Phytochemistry}. Elsevier Science, 2004, vol.~65, No~11, p.~1499-1506. ISSN~0031-9422.

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