Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Elastase activity assay.

MG Manuel R. Gonzalez
BF Betty Fleuchot
LL Leonardo Lauciello
PJ Paris Jafari
LA Lee Ann Applegate
WR Wassim Raffoul
YQ Yok-Ai Que
KP Karl Perron
request Request a Protocol
ask Ask a question
Favorite

Measurement of elastase activity was performed according to the elastin Congo red (ECR) assay previously described in reference 34. Briefly, 10 µl of culture supernatant was mixed with 190 µl of ECR buffer composed of Tris-HCl (100 mM), CaCl2 (1 mM, pH 7.5), and 20 mg/ml of elastin Congo red (Sigma). After agitation 1 h at 37°C, the insoluble ECR was removed by centrifugation (5 min, 2,500 × g). Absorbance of the supernatant was measured at 495 nm by spectrophotometry. Data were normalized to the control background prior normalization to the OD values. Analyses were performed three times in three biological replicates, and standard deviations (and error bars) are indicated.

Copyright and License information:  ©2016 Gonzalez et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A