Stains-All is a Dye that Probes the Conformational Features of Calcium Binding Proteins

  • Yogendra Sharma
  • Doraivajan Balasubramanian


The field of calcium-binding proteins (CaBP) is still growing rapidly with increasing discoveries of these proteins in a variety of tissues. Significant methodological progress has been made in identifying calcium-binding proteins (CaBP) and in investigating their physicochemical and biochemical properties. The usual methods for the detection and identification of calcium-binding proteins used are ultrafiltration, 45Ca overlay in gel electrophoresis and subsequent autoradiography, Chelex competitive calcium binding assay, and use of lanthanum luminescence, particularly of terbium, as calcium mimic. Analysis of calcium binding properties implies various methods such as equilibrium dialysis or flow dialysis, equilibrium gel filtration, spectroscopic procedures, and others (for a review, see Schachtele and Marme 1988). Considering the general interest in calcium and CaBP, we review here the less described but widely used technique of staining using the cationic dye called Stains-all.


Calcium Binding Malignant Hyperthermia Polyaspartic Acid Cardiac Sarcoplasmic Reticulum Sarcoplasmic Reticulum Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Badar JP, Ray DA, Steck TL (1972) Electrophoretic determination of hyaluronate produced by cells in culture. Biochim Biophys Acta 264:73–84.CrossRefGoogle Scholar
  2. Beans RC, Shepherd WC, Kay RE, Walwick ER (1965) Spectral changes in a cationic dye to interaction with macromolecules. III. Stoichiometry and mechanism of the complexing reaction. J Phys Chem 69:43684379.Google Scholar
  3. Caday CG, Steiner RF (1985) The interaction of calmodulin with the carbocyanine dye (Stains-all). J Biol Chem 260:5985–5990.PubMedGoogle Scholar
  4. Caday CG, Lambooy PK, Steiner RF (1986) The interaction of calcium binding proteins with the carbocyanine dye Stains-all. Biopolymers 25:1579–1595.PubMedCrossRefGoogle Scholar
  5. Caday CG, Steiner RF (1986) The interaction of calmodulin with melittin. Biochem Biophys Res Comm 135:419–425.PubMedCrossRefGoogle Scholar
  6. Campbell KP, Maclennan DH, Jorgensen AO (1983) Staining of the calcium binding proteins, calsequestrin, calmodulin, troponin C and S-100 with the cationic dye “Stains-all”. J Biol Chem 258:11267–11273.PubMedGoogle Scholar
  7. Dahlberg AE, Dandman CW, Peacock AC (1969) Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol 41:139–147.PubMedCrossRefGoogle Scholar
  8. Ervasti JM, Ciaessens MT, Mickelson JR, Louis CF (1989) Altered transverse tubule dihydropyridine receptor binding in malignant hyperthermia. J Biol Chem 264:2711–2717.PubMedGoogle Scholar
  9. Fasman GD (1967) Poly α-amino acids. Marcel Dekker, New York.Google Scholar
  10. Green MR, Pastewka JV (1974a) Simultaneous differential staining by a cationic carbocyanine dye of nucleic acids, proteins and conjugated proteins. I. Phosphoproteins. J Histochem Cytochem 22:767–773.PubMedCrossRefGoogle Scholar
  11. Green MR, Pastewka JV (1974b) Simultaneous differential staining by a cationic carbocyanine dye of nucleic acids, proteins and conjugated proteins. II. Carbohydrate and sulfated carbohydrate-containing proteins. J Histochem Cytochem 22:774–781.PubMedCrossRefGoogle Scholar
  12. Green MR, Pastewka JV (1975) Identification of sialic acid rich proteins on polyacrylamide gel. Anal Biochem 65:66–72.PubMedCrossRefGoogle Scholar
  13. Jelley EE (1936) Spectral absorption and fluorescence of dyes in the molecular state. Nature 138:1009–1010.CrossRefGoogle Scholar
  14. Jones LR, Besch HR Jr, Fleming JW, McConnaughley MM, Watanabe AM (1979) Separation of vesicles of cardiac sarcoplasmic reticulum: comparative biochemical analysis of component activities. J Biol Chem 254:530–539.PubMedGoogle Scholar
  15. Jones LR, Cala SE (1981) Biochemical evidence for functional heterogeneity of cardiac sarcoplasmic reticulum vesicles. J Biol Chem 256:11809–11818.PubMedGoogle Scholar
  16. Hofman SL, Brown MS, Lee E, Pathak RK, Anderson RG, Goldstein JL (1989) Purification of a sarcoplasmic reticulum protein that binds calcium and plasma lipoproteins. J Biol Chem 264:8260–8270.Google Scholar
  17. Kay RE, Walwick ER, Gifford CK (1964) Spectral changes in a cationic dye to interaction with macromolecules. I. Behavior of dye alone in solution and the effect of added macromolecules. J Phys Chem 68:1896–1906.CrossRefGoogle Scholar
  18. Kay RE, Walwick ER, Gifford CK (1964) Spectral changes in a cationic dye due to interaction with macromolecules. II. Effects of environment and macromolecule structure. J. Phys Chem 68:1907–1916.CrossRefGoogle Scholar
  19. King LE, Morrison M (1976) The visualization of human erythrocyte membrane proteins and glycoproteins and SDS-polyacrylamide gels employing a single staining procedure. Anal Biochem 71:223–230.PubMedCrossRefGoogle Scholar
  20. Krause KH, Chou M, Thomas MA, Sjolund RD, Campbell KP (1989) Plant cells containing calsequestrin. J Biol Chem 264:4269–4272.PubMedGoogle Scholar
  21. Ksiezak-Reding H, Yen SH (1987) Phosphatase and carbocyanine dye binding define different types of phosphate groups in mammalian neurofilaments. J Neurosci 3554-3560.Google Scholar
  22. Maruyama K, Nonomura Y (1984) High molecular weight protein in the microsome of seallop striated muscle. J Biochem (Tokyo) 96:859–870.Google Scholar
  23. Moore BW (1988) Conformational and hydrophobic properties of rat and bovine S-100 proteins. Neurochem Res 13:539–545.PubMedCrossRefGoogle Scholar
  24. Oberdorf JA, Lebeche D, Head JF, Kaminer B (1988) Identification of a calsequestrin like protein from sea urchin eggs. J Biol Chem 263:6806–6809.PubMedGoogle Scholar
  25. Schachtele CN, Marme D (1988) Methods of assay of calcium binding proteins. In: Calcium-binding proteins, Vol I, (Thompson MP ed.), CRC Press Inc., Florida, pp 83–96.Google Scholar
  26. Sharma Y, Rao CM, Narasu ML, Rao SC, Somasundaram T, Krishna AG, Balasubramanian D (1989a) Calcium ion binding to ß-and to δ-crystallins. The presence of EF-hand motif in δ-crystallin that aids to calcium binding. J Biol Chem 264:12794–12799.PubMedGoogle Scholar
  27. Sharma Y, Rao CM, Rao SC, Krishna AG, Somasundaram T, Balasubramanian D (1989b) Binding site confirmation dictates the color of the dye Stains-all. A study of the binding of this dye to the eye lens proteins crystallins J Biol Chem 264:20923–20927.PubMedGoogle Scholar
  28. Vilanova M, Vendrell J, Cuchillo CM, Ailes FX (1988) Analysis of the conformation and ligand binding properties of the activation segment of pig procarboxypeptidase A. Biochem J 251:901–905.PubMedGoogle Scholar
  29. Wallace RA, Morgan JP (1986a) Chromatographic resolution of chicken phosphovitin. Multiple macromolecular species in a classic vitellogenin-derived phosphoprotein. Biochem J 240:871–878.PubMedGoogle Scholar
  30. Wuytack F, Raeymaekers L, Verbist J, Jones LR, Casteels R (1987) Smooth-muscle endoplasmic reticulum contains a cardiac-like form of calsequestrin. Biochem Biophys Acta 899:151–158.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1991

Authors and Affiliations

  • Yogendra Sharma
  • Doraivajan Balasubramanian

There are no affiliations available

Personalised recommendations