Staining of the Ca2+-binding proteins, calsequestrin, calmodulin troponin C, and S-100, with the cationic carbocyanine dye «Stains-all

Kevin P Campbell; David H MacLennan; Annelise O Jorgensen

doi:10.1016/S0021-9258(17)44413-9

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Staining of the Ca2+-binding proteins, calsequestrin, calmodulin troponin C, and S-100, with the cationic carbocyanine dye «Stains-all

Journal article

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Staining of the Ca2+-binding proteins, calsequestrin, calmodulin troponin C, and S-100, with the cationic carbocyanine dye «Stains-all

Kevin P Campbell, David H MacLennan and Annelise O Jorgensen

The Journal of biological chemistry, Vol.258(18), pp.11267-11273

1983

DOI: 10.1016/S0021-9258(17)44413-9

PMID: 6193121

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Abstract

The Ca2+-binding proteins, calsequestrin, calmodulin, troponin C, and S-100, have all been shown to stain dark blue or purple with the cationic carbocyanine dye “Stains-all”, while most proteins stain red or pink. Stains-all staining of these Ca2+-binding proteins was 50-100% more intense than Coomassie blue staining, and the blue stained calsequestrin and troponin C could be detected in muscle extracts. Densitometric scans of Stains-all-stained gels revealed that interaction of the dye with Ca2+-binding proteins changed the absorption spectrum of the dye. The dye-protein complex absorbed maximally at 615 nm, reflecting a state that results from the binding of individual dye molecules at anionic sites. Stains-all also interacted with undenatured Ca2+-binding proteins in aqueous solution forming a complex absorbing maximally at 600 nm. A simple assay for Ca2+-binding proteins has been developed using this property. These results suggest that the interaction of the dye with anionic sites within these Ca2+-binding proteins produces the dye-protein complex which absorbs at 600-615 nm. Stains-all staining of these Ca2+-binding proteins will be useful in their identification and purification from various tissues. It might also be a valuable tool in the identification of potential Ca2+-binding proteins.

Fundamental and applied biological sciences. Psychology

Proteins

Binding and carrier proteins

Biological and medical sciences

Analytical, structural and metabolic biochemistry

Details

Title: Subtitle: Staining of the Ca2+-binding proteins, calsequestrin, calmodulin troponin C, and S-100, with the cationic carbocyanine dye «Stains-all
Creators: Kevin P Campbell - Univ. Iowa, dep. physiology biophysics
David H MacLennan - Univ. Iowa, dep. physiology biophysics
Annelise O Jorgensen - Univ. Iowa, dep. physiology biophysics
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.258(18), pp.11267-11273
DOI: 10.1016/S0021-9258(17)44413-9
PMID: 6193121
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology; Bethesda, MD
Language: English
Date published: 1983
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9984068262402771

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