Biotin-Labeled Potato Chymotrypsin Inhibitor-1: A Useful Probe for the Detection and Quantitation of Chymotrypsin-like Serine Proteinases on Western Blots and Its Application in the Detection of a Serine Proteinase Synthesized by Articular Chondrocytes
1995
Rodgers, KJ | Melrose, J | Ghosh, P
Potato chymotrypsin inhibitor-1 (pCTI-1) was biotinylated by reaction with sulfosuccinimidyl-6-(biotinamido)hexanoate. This derivative was used as a probe on Western blots for the detection and quantitation of chymotrypsin and the detection of a chymotrypsin-like serine proteinase synthesized by ovine chondrocytes in alginate bead culture. Densitometric analysis demonstrated that there was a linear relationship between the amount of chymotrypsin electrophoresed, over the range 0.1 to 10 ng, and the intensity of the band detected on Western blots using biotinylated pCTI-1 as probe, indicating that the technique could be used for the quantification of active proteinases. The biotinylated pCTI-1 detection technique was convenient to use, reproducible, and more sensitive than zymography. © 1995 Academic Press. All rights reserved.
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