Electrophoretic testing for collagenases (Novex
Zymogram Gel) and measurement of urine elastase activity produced results within the reference intervals (data not shown).
For the interpretation of
zymogram patterns we used the quartenary structure of the enzymes known from other species (Evans 1987; Baker et al.
The study of proteases was complemented by
zymogram using SDS-PAGE electrophoresis and the use of the same inhibitors.
For both zymography and reverse zymography, gels were electrophoresed in 1X
zymogram running buffer (25 mM Tris, 192 mM glycine, and 0.1% SDS), followed by ingel renaturation for 30 min at room temperature in renaturing solution (2.5%Triton X-100).
sanguineus after water exchanged via dialysis were separated by Native-PAGE technique to observe activity
zymogram and protein pattern.
The casein
zymogram analysis (Figure 2B) revealed the presence of a single active band of molecular mass corresponding to that observed in silver-stained SDS-PAGE.
MMPs in the medium released from CL1-0 cells were assayed using gelatin zymography (8%
zymogram gelatin gels) according to the methods reported by Huang et al.
Zymogram of adult and prepubertal quail testes homogenate revealed only single band at identical position when subjected to LDH enzyme specific stain (Figure 2).
The multiple forms of the enzyme activity were detected on
zymogram after non-denaturing PAGE.
Electrophoretic separation and enzyme
zymogram in isoelectrofocusing (IEF gels)
The
zymogram shows bands that are unique to particular species.