96-well plate-based method for total collagen analysis of cell cultures.
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.
Fenwick
S A
SA
Rheumatology Research Unit, Box 194, E6, Addenbrookes Hospital NHS Trust, Hills Road, Cambridge CB2 2QQ, UK. saf25@medschl.cam.ac.uk
Curry
V
V
Clements
S
S
Hazleman
B L
BL
Riley
G P
GP
D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis* / Q000096:biosynthesis; D017364:Collagenases / Q000378:metabolism; D005374:Filtration / Q000295:instrumentation; D006909:Hydroxyproline / Q000032:analysis; D007553:Isotope Labeling; D011392:Proline / Q000378:metabolism; D012588:Scintillation Counting / Q000295:instrumentation; D014316:Tritium
D002250:Carbon Radioisotopes; D014316:Tritium; D003094:Collagen; D011392:Proline; D017364:Collagenases; D006909:Hydroxyproline