TY - JOUR
T1 - Bromobimane probes for thiols
AU - Kosower, Edward M.
AU - Kosower, Nechama S.
N1 - Funding Information:
The authors are grateful for the support of their research on thiols and thiol agents by the National Institutes of Health, the United States-Israel Binational Science Foundation, the Israel Academy of Sciences, the Chief Scientist's Office, Israel Ministry of Health, the European Research Office, the Petroleum Research Fund, the J. S. Guggenheim Foundation, and the National Science Foundation.
Funding Information:
The authors are grateful for the support of their research on thiol and thiol agents by the National Institutes of Health, the United States-Israel Binational Science Foundation, the Israel Academy of Science, the Chief Scientist's Office, Israel Ministry of Health, the European Research Office, the Petroleum Research Fund, the J. S. Gnggenheim Foundation, the Rockefeller Foundation, and the National Science Foundation.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - This chapter discusses bromobimane probes for thiols and describes the use of four bromobimanes for fluorescent labeling of biochemical and biological systems. The four bromobimanes are (1) mBBr, (2) bBBr, (3) qBBr, and (4) SBBr. The bromobimanes are essentially nonfluorescent and are relatively stable in the dry state when stored in the dark. Chromatography on thin-layer silica yields a yellow nonfluorescent spot that develops a blue fluorescence after several minutes of exposure to 360-nm light. The change is a convenient characteristic for the identification of the bromobimanes and suggests sensitivity to light (photolysis). Bromobimanes in solution react with small thiols (e.g., the tripeptide thiol glutathione [GSH]), and with reactive protein thiol groups (e.g., hemoglobin). The reactions of bromobimanes with thiols are second order and dependent on pH, the active nucleophile being the thiolate anion, such as GS−. The reaction of bromobimane with a thiolate converts the nonfluorescent agent into water-soluble fluorescent products. The chapter discusses the chemical and photophysical properties of bromobimanes, labeling procedures, labeling of thiols in tissues, and other applications of thiol labeling.
AB - This chapter discusses bromobimane probes for thiols and describes the use of four bromobimanes for fluorescent labeling of biochemical and biological systems. The four bromobimanes are (1) mBBr, (2) bBBr, (3) qBBr, and (4) SBBr. The bromobimanes are essentially nonfluorescent and are relatively stable in the dry state when stored in the dark. Chromatography on thin-layer silica yields a yellow nonfluorescent spot that develops a blue fluorescence after several minutes of exposure to 360-nm light. The change is a convenient characteristic for the identification of the bromobimanes and suggests sensitivity to light (photolysis). Bromobimanes in solution react with small thiols (e.g., the tripeptide thiol glutathione [GSH]), and with reactive protein thiol groups (e.g., hemoglobin). The reactions of bromobimanes with thiols are second order and dependent on pH, the active nucleophile being the thiolate anion, such as GS−. The reaction of bromobimane with a thiolate converts the nonfluorescent agent into water-soluble fluorescent products. The chapter discusses the chemical and photophysical properties of bromobimanes, labeling procedures, labeling of thiols in tissues, and other applications of thiol labeling.
UR - http://www.scopus.com/inward/record.url?scp=0029061415&partnerID=8YFLogxK
U2 - 10.1016/0076-6879(95)51117-2
DO - 10.1016/0076-6879(95)51117-2
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AN - SCOPUS:0029061415
SN - 0076-6879
VL - 251
SP - 133
EP - 148
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -