Microplate reader-based quantitation of collagens.
By using a picrosirius dye, sensitive and specific staining of collagens plated in microtiter wells was achieved. The range of detection was from 0.5 to 20 micrograms. Human collagen types I, III, IV, and V were tested and able to be detected by the method. The dye did not bind to acetylcholinesterase or elastin. It did bind to C1q to some extent but this is not surprising since the molecule contains some triple helical collagen-like structures. A comparison performed between this assay and a colorimetric assay for hydroxyproline using tissue culture supernatants gave similar results for both samples. Due to its simplicity and sensitivity this assay will be most useful in laboratories where large numbers of samples must be screened for collagen production.
Walsh
B J
BJ
Centre for Immunology St. Vincent's Hospital, Sydney, New South Wales, Australia.
Thornton
S C
SC
Penny
R
R
Breit
S N
SN
D000110:Acetylcholinesterase / Q000378:metabolism; D001391:Azo Compounds / Q000378:metabolism; D002460:Cell Line; D003094:Collagen / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D003124:Colorimetry; D004396:Coloring Agents / Q000378:metabolism; D004549:Elastin / Q000378:metabolism; D006801:Humans; D006909:Hydroxyproline / Q000032:analysis*; D010851:Picrates / Q000378:metabolism; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D001391:Azo Compounds; D004396:Coloring Agents; D010851:Picrates; C009798:C.I. direct red 80; D003094:Collagen; D004549:Elastin; D000110:Acetylcholinesterase; D006909:Hydroxyproline