Abstract
Gelatinase zymography is a commonly used laboratory procedure; however, variability in sample loading and concentration reduce the accuracy of quantitative results obtained from this technique. To facilitate normalization of gelatinase activity by loaded protein amount, we developed a protocol using the trihalocompound 2,2,2-trichloroethanol to allow for gelatin zymography and total protein labeling within the same gel. We showed that detected protein levels increased linearly with loading, and describe a loading concentration range over which normalized gelatinase activity was constant. We conclude that in-gel total protein detection is feasible in gelatin zymography and greatly improves comparison of gelatinase activity between samples.
Original language | English |
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Pages (from-to) | 8-10 |
Number of pages | 3 |
Journal | Analytical Biochemistry |
Volume | 521 |
DOIs | |
Publication status | Published - 15 Mar 2017 |
Externally published | Yes |
Keywords
- Gelatinase
- Loading controls
- Matrix metalloproteinases
- Trihalocompounds
- Zymography