Regular ArticleCalibrating Gelatin Zymograms with Human Gelatinase Standards
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2012, Clinical BiochemistryCitation Excerpt :In an effort to identify proteases in these biofluids, the zymogram bands were digested with trypsin and analyzed by LC-MS/MS. The proteases identified in each fluid, either by mass spectrometry or western blot analysis, are presented in Table 1. As expected based on zymo incubated with EDTA, western blot and on literature [20–22], several metalloproteinases were identified in all fluids analyzed, namely MMP-2, MMP-9 and pro-MMP-9. These proteases were only identified by western blotting.
Classically activated macrophages use stable microtubules for matrix metalloproteinase-9 (MMP-9) secretion
2012, Journal of Biological ChemistryCitation Excerpt :The absence of the active 82-kDa form in the cell culture supernatant may be a result of culture conditions, in which MMP-9 is diluted away from its activators in the medium (22). Although gelatin zymography is a common tool used to assess the levels of gelatinases (32), to measure the amount of secreted and intracellular MMP-9 protein levels, we next turned to immunoblotting. Western blot analysis of MMP-9 in the total cell lysate consistently revealed two bands of apparent molecular masses of 85 and 92 kDa (Fig. 1C).
Modifications of the defense and remodeling functionalities of bovine neutrophils inside the mammary gland of milk stasis cows received a commercial dry-cow treatment
2011, Veterinary Immunology and ImmunopathologyCitation Excerpt :MMP 9 level was expressed as relative terms to that of incubation without skimmed d 7-dry secretion (0 μg/ml), or to that before incubation (0 min). Samples of human plasma, bovine plasma, bovine skimmed milk serum, and bovine milk somatic cell lysate were also analyzed by gelatin-zymography at regular intervals to serve as references of different gelatinase subtypes (Makowski and Ramsby, 1996; Weng et al., 2008). Human plasma was ethically prepared from the venous blood of a clinically healthy young male.
Assays for determination of matrix metalloproteinases and their activity
2011, TrAC - Trends in Analytical ChemistryCitation Excerpt :Absence of a universal standard contributes to the lack of inter-laboratory reproducibility and transferability of results [38]. The best established standard is human capillary blood due to its availability [39]. Despite the problems with standardization, this method is widely used in tissue-sample analysis [38,40].
Extracellular matrix roles during cardiac repair
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