AN INDIRECT IMMUNOFLUORESCENCE DOUBLE STAINING PROCEDURE FOR THE SIMULTANEOUS FLOW CYTOMETRIC MEASUREMENT OF IODODEOXYURIDINE AND CHLORODEOXYURIDINE INCORPORATED INTO DNA

被引：27

作者：

BAKKER, PJM ^{[1
]}

STAP, J ^{[1
]}

TUKKER, CJ ^{[1
]}

VANOVEN, CH ^{[1
]}

VEENHOF, CHN ^{[1
]}

ATEN, J ^{[1
]}

机构：

[1] UNIV AMSTERDAM,ACAD MED CTR,RADIOBIOL LAB,1105 AZ AMSTERDAM,NETHERLANDS

来源：

CYTOMETRY | 1991年 / 12卷 / 04期

关键词：

IMMUNOCHEMISTRY; DOUBLE STAINING PROCEDURE; IDURD; CLDURD; FLOW CYTOMETRY; CELL KINETICS;

D O I：

10.1002/cyto.990120412

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In this paper we describe an indirect fluorescence double staining procedure for the simultaneous detection of IdUrd and CldUrd in the same cell nucleus. Two commercially available antibodies were selected for this purpose. A rat anti-BrdUrd monoclonal antibody from Seralab was found to bind specifically to CldUrd and BrdUrd. A mouse monoclonal anti-BrdUrd antibody from Becton Dickinson used in a 1:2 dilution binds to all halogenated deoxyuridines but, when the cells were extensively washed with Tris buffer with a high salt concentration, almost no binding to CldUrd was observed. An immunofluorescence procedure was developed, based on these primary antibodies, raised in different species (rat and mouse), in combination with highly purified second antibodies: FITC conjugated goat antirat and Texas-Red conjugated goat antimouse.

引用

页码：366 / 372

页数：7

共 18 条

[1] FLOW CYTOMETRIC ANALYSIS OF EXPERIMENTAL PARAMETERS FOR THE IMMUNOFLUORESCENT LABELING OF BRDURD IN VARIOUS TUMOR-CELL LINES [J].

BAKKER, PJM ;

ATEN, JA ;

TUKKER, CJ ;

BARENDSEN, GW ;

VEENHOF, CHN .

HISTOCHEMISTRY, 1989, 91 (05) :425-429

[2] HUMAN-TUMOR CELL-KINETICS USING A MONOCLONAL-ANTIBODY AGAINST IODODEOXYURIDINE - INTRATUMOR SAMPLING VARIATIONS [J].

BEGG, AC ;

MOONEN, L ;

HOFLAND, I ;

DESSING, M ;

BARTELINK, H .

RADIOTHERAPY AND ONCOLOGY, 1988, 11 (04) :337-347

[3] A METHOD TO MEASURE THE DURATION OF DNA-SYNTHESIS AND THE POTENTIAL DOUBLING TIME FROM A SINGLE SAMPLE [J].

BEGG, AC ;

MCNALLY, NJ ;

SHRIEVE, DC ;

KARCHER, H .

CYTOMETRY, 1985, 6 (06) :620-626

[4] AN IMPROVED IMMUNOCYTOCHEMICAL PROCEDURE FOR HIGH-SENSITIVITY DETECTION OF INCORPORATED BROMODEOXYURIDINE [J].

BEISKER, W ;

DOLBEARE, F ;

GRAY, JW .

CYTOMETRY, 1987, 8 (02) :235-239

[5] FLOW CYTOMETRIC MEASUREMENT OF TOTAL DNA CONTENT AND INCORPORATED BROMODEOXYURIDINE [J].

DOLBEARE, F ;

GRATZNER, H ;

PALLAVICINI, MG ;

GRAY, JW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (18) :5573-5577

[6] CYTO-CHEMISTRY FOR BROMODEOXYURIDINE DNA ANALYSIS - STOICHIOMETRY AND SENSITIVITY [J].

DOLBEARE, F ;

BEISKER, W ;

PALLAVICINI, MG ;

VANDERLAAN, M ;

GRAY, JW .

CYTOMETRY, 1985, 6 (06) :521-530

[7]

DOLBEARE F, 1988, P AM ASSOC CANC RES, V29, P1896

[8] MONOCLONAL-ANTIBODY TO 5-BROMODEOXYURIDINE AND 5-IODODEOXYURIDINE - A NEW REAGENT FOR DETECTION OF DNA-REPLICATION [J].

GRATZNER, HG .

SCIENCE, 1982, 218 (4571) :474-475

[9] S-PHASE FRACTION OF HUMAN-BRAIN TUMORS INSITU MEASURED BY UPTAKE OF BROMODEOXYURIDINE [J].

HOSHINO, T ;

NAGASHIMA, T ;

CHO, KG ;

MUROVIC, JA ;

HODES, JE ;

WILSON, CB ;

EDWARDS, MSB ;

PITTS, LH .

INTERNATIONAL JOURNAL OF CANCER, 1986, 38 (03) :369-374

[10]

KARCHER H, 1987, STRAHLENTHER ONKOL, V163, P195

← 1 2 →