+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

An automated micro-fluorometric assay for monitoring oxidative burst activity of phagocytes



An automated micro-fluorometric assay for monitoring oxidative burst activity of phagocytes



Journal of Immunological Methods 159(1-2): 131-138



A micro-fluorometric assay using 2',7'-dichlorofluorescin diacetate (DCFH-DA) to monitor oxidative burst (OB) in phagocytes has been developed. This assay is based on the oxidation of nonfluorescent DCFH-DA to highly fluorescent 2',7'-dichlorofluorescein (DCF) both intracellularly and extracellularly. A murine macrophage cell line, J774, and a human monocytic cell line, Mono Mac 6, were used as models. The cells were harvested from tissue culture flasks, washed, counted and adjusted to desired concentrations. They were then dispensed into a 96-well flat-bottom tissue culture plate. After adding DCFH-DA and a agent eliciting OB, the plates were incubated in 5% CO-2 at 37 degree C for various periods. The intensity of fluorescence was measured directly in the wells of the tissue culture plate with the cells in situ using a computerized microplate fluorometer at 485 nm excitation and 530 nm emission. This assay provided a rapid measurement of oxidative burst of phagocytes. The automated micro-fluorometric assay may be suitable for screening the immunomodulating activities of various biological and pharmacological substances.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 008142184

Download citation: RISBibTeXText

PMID: 8445246

DOI: 10.1016/0022-1759(93)90150-6


Related references

Flow cytometric assays of oxidative burst activity in phagocytes. Methods in Enzymology 233: 539-548, 1994

Automated micro-fluorometric assays for monitoring phagocyte functions. Abstracts of the General Meeting of the American Society for Microbiology 93: 555, 1993

New automated fluorometric peptide micro assay for carnosine in mouse olfactory bulb. Analytical Biochemistry 86(2): 670-678, 1978

Changes in the antioxidative capacity of the plasma, the oxidative burst and phagocytic activity of phagocytes from patients with ALD. Alcoholism Clinical and Experimental Research 24(5 Suppl.): 199A, 2000

Automated image analysis for monitoring oxidative burst in macrophages. Analytical and Quantitative Cytology and Histology 22(5): 423-427, 2000

Immunoenhancing effect of 1,2-benzopyrone on the oxidative burst activity to phagocytic response of canine peripheral blood phagocytes. Journal of Veterinary Clinics 21(3): 236-242, 2004

Analysis of lipase activity in duodenal juice. Comparison of an automated spectrophotometric assay to a fluorometric microplate assay, and factors affecting sample stability. Scandinavian Journal of Gastroenterology 53(10-11): 1206-1211, 2018

Influence of fluconazole on phagocytosis, oxidative burst and killing activity of human phagocytes. Using a flow cytometric method with whole blood. European Journal of Medical Research 5(10): 455-459, 2000

An automated fluorometric assay for proteolytic activity in wheat. Cereal Chemistry 52(4): 451-458, 1975

Effects of milking frequency on phagocytosis and oxidative burst activity of phagocytes from primiparous and multiparous dairy cows during early lactation. Journal of Dairy Science 91(2): 587-595, 2008

Oxidative burst activity of cerebrospinal fluid phagocytes and neuronal damage are reduced by rifampin in experimental Streptococcus pneumoniae meningitis. Neurology 54(7 Supp 3): A288, 2000

Immunoenhancing effect of trans-10, cis-12 conjugated linoleic acid on the phagocytic capacity and oxidative burst activity of canine peripheral blood phagocytes. Research in Veterinary Science 85(2): 269-278, 2008

Detection of D-arabinitol in serum by rapid automated enzymatic fluorometric assay Diagnosis and therapeutic monitoring of invasive candidiasis. Abstracts of the Interscience Conference on Antimicrobial Agents and Chemotherapy 40: 378, 2000

Pectolinarigenin from the leaves of Clerodendrum volubile shows potent immunomodulatory activity by inhibiting T - cell proliferation and modulating respiratory oxidative burst in phagocytes. Biomedicine and PharmacoTherapy 93: 529-535, 2017

Automated fluorometric creatine kinase assay. Measurement of 100-fold normal activity without serum dilution. Clinical Chemistry 20(5): 560-565, 1974