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96-well plate-based method for total collagen analysis of cell cultures
S A Fenwick1, V Curry, S Clements
1Rheumatology Research Unit, Box 194, E6, Addenbrookes Hospital NHS Trust, Hills Road, Cambridge CB2 2QQ, UK. saf25@medschl.cam.ac.uk
Abstract:
Total collagen assays are often laborious and use large quantities of consumables. We have developed a new method of assaying total 3H-proline-labeled collagen from cultured cells. Cells and media are harvested from 96-well plates directly onto fiberglass filtermats and counted in the Wallac 1205 flat-bed scintillation counter (BetaPlate). The assay was validated by comparison with a traditional total collagen assay. The resulting assay provides a rapid one-step method for quantifying collagen synthesis, which, unlike many collagen assays, does not require extensive dialysis or precipitation of proteins.