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 16 条

[1] FLOW CYTOMETRIC MEASUREMENT OF TOTAL DNA CONTENT AND INCORPORATED BROMODEOXYURIDINE
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
[2] A NEW DUAL STAINING TECHNIQUE FOR SIMULTANEOUS FLOW CYTOMETRIC DNA ANALYSIS OF LIVING AND DEAD CELLS
STOHR, M
VOGTSCHADEN, M
ACTA PATHOLOGICA ET MICROBIOLOGICA SCANDINAVICA SECTION A-PATHOLOGY, 1981, : 96 - 99
[3] SIMULTANEOUS FLOW CYTOMETRIC DETECTION OF CELLULAR C-MYC PROTEIN, INCORPORATED BROMODEOXYURIDINE, AND DNA
ROSETTE, CD
DETERESA, PS
PALLAVICINI, MG
CYTOMETRY, 1990, 11 (04): : 547 - 551
[4] FLOW CYTOMETRIC DETECTION OF HERPES-SIMPLEX VIRUS TYPE-2 INFECTED AND TRANSFORMED-CELLS BY IMMUNOENZYMATIC AND BY INDIRECT IMMUNOFLUORESCENCE STAINING
GOOLSBY, C
GAY, H
DOCHERTY, JJ
TODD, P
CYTOMETRY, 1988, 9 (02): : 126 - 130
[5] RELIABILITY OF DNA MEASUREMENT AFTER SIMULTANEOUS STAINING OF DNA AND RNA BY ACRIDINE-ORANGE FOR FLOW-CYTOPHOTOMETRY
ZITTOUN, R
POCHAT, L
MATHIEU, M
POCHAT, R
PATHOLOGIE BIOLOGIE, 1981, 29 (05): : 317 - 320
[6] SIMULTANEOUS DEMONSTRATION OF MULTIPLE ANTIGENS BY INDIRECT IMMUNOFLUORESCENCE OR IMMUNOGOLD STAINING - NOVEL LIGHT AND ELECTRON MICROSCOPICAL DOUBLE AND TRIPLE STAINING METHOD EMPLOYING PRIMARY ANTIBODIES FROM THE SAME SPECIES
WANG, BL
LARSSON, LI
HISTOCHEMISTRY, 1985, 83 (01) : 47 - 56
[7] WASHLESS DOUBLE STAINING OF UNFIXED NUCLEI FOR FLOW CYTOMETRIC ANALYSIS OF DNA AND A NUCLEAR ANTIGEN (KI-67 OR BROMODEOXYURIDINE)
LARSEN, JK
CHRISTENSEN, IJ
CHRISTIANSEN, J
MORTENSEN, BT
CYTOMETRY, 1991, 12 (05): : 429 - 437
[8] ONE-STEP PROPIDIUM IODIDE (PI) STAINING FAILURE FOR FLOW CYTOMETRIC DNA CONTENT MEASUREMENT IN MICE LIVER
VAGO, P
AGUILAR, V
ABOUDKHIL, S
GARRELLY, L
BUREAU, JP
BASIC AND APPLIED HISTOCHEMISTRY, 1988, 32 (03): : 394 - 394
[9] Flow acetone-staining technique: A highly efficient procedure for the simultaneous analysis of DNA content, cell morphology, and immunophenotype by flow cytometry
Carbonari, Maurizio
Mancaniello, Debora
Tedesco, Tiziana
Fiorilli, Massimo
CYTOMETRY PART A, 2008, 73A (02) : 168 - 174
[10] FLOW CYTOMETRIC 2-COLOR STAINING TECHNIQUE FOR SIMULTANEOUS DETERMINATION OF HUMAN ERYTHROCYTE-MEMBRANE ANTIGEN AND INTRACELLULAR MALARIAL DNA
PATTANAPANYASAT, K
WEBSTER, HK
UDOMSANGPETCH, R
WANACHIWANAWIN, W
YONGVANITCHIT, K
CYTOMETRY, 1992, 13 (02): : 182 - 187

← 1 2 →