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Staining of the Ca2+-binding Proteins, Calsequestrin, Calmodulin, Troponin C, and S-100, With the Cationic Carbocyanine Dye "Stains-all"

Staining of the Ca2+-binding Proteins, Calsequestrin, Calmodulin, Troponin C, and S-100, With the Cationic Carbocyanine Dye "Stains-all"

K P Campbell, D H MacLennan, A O Jorgensen

J Biol Chem. 1983 Sep 25;258(18):11267-73.

PMID: 6193121

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP7423316	Stains-All		7423-31-6	Price

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