azaguanine and Inflammation

This protocol describes microsphere-based protease assays for use in flow cytometry and high-throughput screening. This platform measures a loss of fluorescence from the surface of a microsphere due to the cleavage of an attached fluorescent protease substrate by a suitable protease enzyme. The assay format can be adapted to any site or protein-specific protease of interest and results can be measured in both real time and as endpoint fluorescence assays on a flow cytometer. Endpoint assays are easily adapted to microplate format for flow cytometry high-throughput analysis and inhibitor screening.

Topics: Animals; Biotinylation; Flow Cytometry; Fluorescence Resonance Energy Transfer; Green Fluorescent Proteins; High-Throughput Screening Assays; Humans; Inflammation; Kinetics; Microspheres; Peptide Hydrolases; Peptides; Reproducibility of Results; Temperature

Topics: Agranulocytosis; Aminopterin; Animals; Azaguanine; Biopsy; Chloramphenicol; Connective Tissue; Cyclophosphamide; Dactinomycin; Fluorouracil; Glycosaminoglycans; Histocytochemistry; Inflammation; Leukopenia; Lymphocytes; Mercaptopurine; Mice; Neutrophils; Photomicrography; Puromycin; Rabbits; Research

Topics: Aminopterin; Antimetabolites; Azaguanine; Chloramphenicol; Connective Tissue; Cyclophosphamide; Dactinomycin; Egg White; Fluorouracil; Inflammation; Leukocyte Count; Lymphocytes; Mice; Pathology; Pharmacology; Proteins; Puromycin; Rabbits; Research; Toxicology