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.
analysis detected a single band of activity in each lane of the gel (Figure 1).
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.
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)
shows bands that are unique to particular species.