DETECTING ENZYMATIC-ACTIVITY IN CELLS USING FLUOROGENIC SUBSTRATES

被引:26
作者
HAUGLAND, RP
机构
[1] Molecular Probes, Inc., Eugene, OR, 97402
关键词
FLUORESCENCE; ENZYME; B-GALACTOSIDASE; LIVE CELLS; HISTOCHEMISTRY; SUBSTRATES;
D O I
10.3109/10520299509108201
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Fluorogenic substrates can detect enzymatic activity associated with cells, It is difficult, however, to detect activity within a single cell or in an organelle since hydrolytic substrates yield products that rapidly leak from the cell. Several new solutions are presented including trapping the fluorescent product in membranes, in cell organelles, or as a glutathione conjugate. Novel substrates also are described that directly yield highly fluorescent precipitates at the site of enzymatic activity, These can be used for detecting endogenous activity in cells or for enzyme-amplified histochemical detection, Some of these substrates can be used in live cells.
引用
收藏
页码:243 / 251
页数:9
相关论文
共 17 条
  • [11] FLOW CYTOMETRIC ANALYSIS OF PROTEASE ACTIVITIES IN VITAL CELLS
    ROTHE, G
    KLINGEL, S
    ASSFALGMACHLEIDT, I
    MACHLEIDT, W
    ZIRKELBACH, C
    BANATI, RB
    MANGEL, WF
    VALET, G
    [J]. BIOLOGICAL CHEMISTRY HOPPE-SEYLER, 1992, 373 (07): : 547 - 554
  • [12] DIHYDRORHODAMINE-123 - A NEW FLOW CYTOMETRIC INDICATOR FOR RESPIRATORY BURST ACTIVITY IN NEUTROPHIL GRANULOCYTES
    ROTHE, G
    OSER, A
    VALET, G
    [J]. NATURWISSENSCHAFTEN, 1988, 75 (07) : 354 - 355
  • [13] MEASUREMENT OF SUPEROXIDE RELEASE IN THE PHAGOVACUOLES OF IMMUNE COMPLEX-STIMULATED HUMAN NEUTROPHILS
    RYAN, TC
    WEIL, GJ
    NEWBURGER, PE
    HAUGLAND, R
    SIMONS, ER
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 1990, 130 (02) : 223 - 233
  • [14] DIHYDROTETRAMETHYLROSAMINE - A LONG WAVELENGTH, FLUOROGENIC PEROXIDASE SUBSTRATE EVALUATED INVITRO AND IN A MODEL PHAGOCYTE
    WHITAKER, JE
    MOORE, PL
    HAUGLAND, RP
    HAUGLAND, RP
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 175 (02) : 387 - 393
  • [15] YANG JW, 1993, BIOTECHNIQUES, V15, P848
  • [16] A NONRADIOACTIVE ASSAY FOR TRANSFECTED CHLORAMPHENICOL ACETYLTRANSFERASE ACTIVITY USING FLUORESCENT SUBSTRATES
    YOUNG, SL
    BARBERA, L
    KAYNARD, AH
    HAUGLAND, RP
    KANG, HC
    BRINKLEY, M
    MELNER, MH
    [J]. ANALYTICAL BIOCHEMISTRY, 1991, 197 (02) : 401 - 407
  • [17] DETECTING LACZ-GENE EXPRESSION IN LIVING CELLS WITH NEW LIPOPHILIC, FLUOROGENIC BETA-GALACTOSIDASE SUBSTRATES
    ZHANG, YZ
    NALEWAY, JJ
    LARISON, KD
    HUANG, ZJ
    HAUGLAND, RP
    [J]. FASEB JOURNAL, 1991, 5 (15) : 3108 - 3113