Tissue Harvest for Protein Gels
Clear arteries well in situ, and after removal rinse and flush with Ringer¡¦s in a petri dish. Blot tissue to dry, and snap freeze in liquid N2. Tissue can be frozen at -70C indefinitely.
1. Fill cooling...
Protocol for Reverse Zymograms
Separating/Reverse Zymo Gel
3.75 ml 4X Tris-Cl, pH 8.8
3.25 ml distilled water
6.50 ml Protogel (30% mix 29:1 acrylamide:bisacrylamide (3.3% C) Biorad 161-0156
1.5 ml gelatin (1% solution)
In Situ Zymography protocol
1. Cut 8 um cryosections of mouse arterial tissue and place on + charged slides as shown below. This arrangement allows for assay of 2 conditions per slide
2. Keep cut slides at -80 °C until use.
3. Wash slides...
Recently we demonstrated that intracellular calcium regulates proliferation of hepatic stellate cells via CaMK II. We now have preliminary evidence that CaMK II regulates matrix release and myofibroblastic differentiation of hepatic stellate cells and...