Western blot analysis is shown using anti-Collagen I
antibody to detect expression of collagen I in Wistar rat
hepatic stellate cells (HSC) in control (GFP-transduced) (left
lane) and PPARg-transduced cell lysates (right lane).
Protein
staining shown below each blot depicts equal protein
loading. An equal amount of the whole cell protein (100 mg)
was separated by SDS-PAGE and electroblotted to
nitrocellulose
membranes. Proteins were detected by
incubating the membrane with anti- Collagen I antibody at a
concentration of 0.2 - 2 mg/10 ml in TBS (100 mM Tris-HCl,
0.15 M NaCl, pH 7.4) with 5% Non-fat milk. Detection
occurred by incubation with a horseradish peroxidaseconjugated
secondary antibody at 1 mg/10 ml. Proteins were
detected by a chemiluminescent method using the PIERCE
ECL kit (Amersham Biosciences). Other detection systems
will yield similar results. See Hazra et al. (2004) for
additional details.