An enzyme-linked immunosorbent assay for bromodeoxyuridine incorporation using fixed microcultures

Journal Article · Thu Mar 01 00:00:00 EST 1990 · Analytical Biochemistry; (USA)

DOI:https://doi.org/10.1016/0003-2697(90)90310-6· OSTI ID:6727537

Muir, D; Varon, S; Manthorpe, M ^[1]

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1. Univ. of California, San Diego, La Jolla (USA)

We report a quantitative method by which a single microculture can be examined for cell morphology; cell number; DNA synthesis; and expression of cell antigens. This method first involves measuring by enzyme-linked immunosorbent assay (ELISA) the total bromodeoxyuridine (BrdU) incorporation into DNA by monolayer microcultures. The BrdU-ELISA measurement was followed by simultaneous immunostaining for BrdU-positive nuclei and for a cytoplasmic antigen. The method was applied to the measurement of mitogen-induced proliferation of rat sciatic nerve Schwann cell and cerebral astroglia microcultures. The ELISA measurement of BrdU incorporation compares favorably with measurements of tritiated thymidine incorporation and offers the additional advantages that the same microculture can subsequently be examined for cell number, for cell morphology, and for the percentage of cells having BrdU-labeled nuclei and other antigens.

• OSTI ID: 6727537
• Journal Information: Analytical Biochemistry; (USA), Vol. 185:2; ISSN 0003-2697
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
BUDR
ENZYME IMMUNOASSAY
NERVE CELLS
CELL PROLIFERATION
ANTIBODIES
ANTIGENS
CELL CULTURES
CELL NUCLEI
CULTURE MEDIA
DNA REPLICATION
GOATS
MITOGENS
MORPHOLOGY
RATS
THYMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMAL CELLS
ANIMALS
ANTIMETABOLITES
AZINES
BIOASSAY
BROMOURACILS
CELL CONSTITUENTS
DOMESTIC ANIMALS
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
IMMUNOASSAY
ISOTOPE APPLICATIONS
MAMMALS
NUCLEIC ACID REPLICATION
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RIBOSIDES
RODENTS
RUMINANTS
SOMATIC CELLS
URACILS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques