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A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion

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A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion. / Tolosa, E; Shaw, S.

in: J IMMUNOL METHODS, Jahrgang 192, Nr. 1-2, 10.06.1996, S. 165-72.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Tolosa, E & Shaw, S 1996, 'A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion', J IMMUNOL METHODS, Jg. 192, Nr. 1-2, S. 165-72.

APA

Tolosa, E., & Shaw, S. (1996). A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion. J IMMUNOL METHODS, 192(1-2), 165-72.

Vancouver

Tolosa E, Shaw S. A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion. J IMMUNOL METHODS. 1996 Jun 10;192(1-2):165-72.

Bibtex

@article{e2f43869ea8343128efcd0797b18df3c,
title = "A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion",
abstract = "Assays of cell adhesion generally require prelabeling of cells with radioactive or fluorescent probes. A new fluorogenic phosphatase assay requiring no prelabeling has been developed to quantitate cell number, which can thus serve as the basis for quantitating cell adhesion or migration. The assay uses the non-fluorescent substrate 3,6-fluorescein diphosphate (FDP) whose dephosphorylation generates fluorescein. The fluorescence generated is linear with incubation time and cell number until substrate becomes limiting; the assay easily quantitates cells over a range from 10(3) to 10(6) for a variety of cell types, including resting T cells. It is as sensitive as the 51Cr assay, but has the many advantages of a non-radioactive assay, making more convenient the removal of nonadherent cells by simple 1 x g sedimentation. Unlike most other non-radioactive assays, it requires no pre-incubation; this: (1) reduces cell manipulation; (2) eliminates problems of spontaneous release; and (3) avoids potential dye toxicity. This technique of cell quantitation has been adopted as standard in our laboratory for routine adhesion and migration assays.",
keywords = "Buffers, Cell Adhesion, Cell Line, Centrifugation, Chromium Radioisotopes, Enzyme Activation, Fluoresceins, Fluorescent Dyes, Humans, Hydrogen-Ion Concentration, Lymphocyte Count, Phosphoric Monoester Hydrolases, T-Lymphocytes, Tumor Cells, Cultured",
author = "E Tolosa and S Shaw",
year = "1996",
month = jun,
day = "10",
language = "English",
volume = "192",
pages = "165--72",
journal = "J IMMUNOL METHODS",
issn = "0022-1759",
publisher = "Elsevier",
number = "1-2",

}

RIS

TY - JOUR

T1 - A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion

AU - Tolosa, E

AU - Shaw, S

PY - 1996/6/10

Y1 - 1996/6/10

N2 - Assays of cell adhesion generally require prelabeling of cells with radioactive or fluorescent probes. A new fluorogenic phosphatase assay requiring no prelabeling has been developed to quantitate cell number, which can thus serve as the basis for quantitating cell adhesion or migration. The assay uses the non-fluorescent substrate 3,6-fluorescein diphosphate (FDP) whose dephosphorylation generates fluorescein. The fluorescence generated is linear with incubation time and cell number until substrate becomes limiting; the assay easily quantitates cells over a range from 10(3) to 10(6) for a variety of cell types, including resting T cells. It is as sensitive as the 51Cr assay, but has the many advantages of a non-radioactive assay, making more convenient the removal of nonadherent cells by simple 1 x g sedimentation. Unlike most other non-radioactive assays, it requires no pre-incubation; this: (1) reduces cell manipulation; (2) eliminates problems of spontaneous release; and (3) avoids potential dye toxicity. This technique of cell quantitation has been adopted as standard in our laboratory for routine adhesion and migration assays.

AB - Assays of cell adhesion generally require prelabeling of cells with radioactive or fluorescent probes. A new fluorogenic phosphatase assay requiring no prelabeling has been developed to quantitate cell number, which can thus serve as the basis for quantitating cell adhesion or migration. The assay uses the non-fluorescent substrate 3,6-fluorescein diphosphate (FDP) whose dephosphorylation generates fluorescein. The fluorescence generated is linear with incubation time and cell number until substrate becomes limiting; the assay easily quantitates cells over a range from 10(3) to 10(6) for a variety of cell types, including resting T cells. It is as sensitive as the 51Cr assay, but has the many advantages of a non-radioactive assay, making more convenient the removal of nonadherent cells by simple 1 x g sedimentation. Unlike most other non-radioactive assays, it requires no pre-incubation; this: (1) reduces cell manipulation; (2) eliminates problems of spontaneous release; and (3) avoids potential dye toxicity. This technique of cell quantitation has been adopted as standard in our laboratory for routine adhesion and migration assays.

KW - Buffers

KW - Cell Adhesion

KW - Cell Line

KW - Centrifugation

KW - Chromium Radioisotopes

KW - Enzyme Activation

KW - Fluoresceins

KW - Fluorescent Dyes

KW - Humans

KW - Hydrogen-Ion Concentration

KW - Lymphocyte Count

KW - Phosphoric Monoester Hydrolases

KW - T-Lymphocytes

KW - Tumor Cells, Cultured

M3 - SCORING: Journal article

C2 - 8699013

VL - 192

SP - 165

EP - 172

JO - J IMMUNOL METHODS

JF - J IMMUNOL METHODS

SN - 0022-1759

IS - 1-2

ER -