Immunohistochemistry of Collagen I antibody. Tissue: Normal Kidney. Fixation: formalin fixed paraffin embedded. Antigen retrieval: No antigen retrieval was performed. Primary antibody: Collagen I 1:100 4 hours at room temperature Secondary antibody: Peroxidase goat anti-rabbit at 1:10000 for 45 min at RT. Localization: Distal tubules in normal kidney tissue. Note the absence of staining of glomeruli. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Anti-Collagen I Antibody - Western Blot. Western blot analysis is shown using Affinity Purified 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 ug) was separated by SDS-PAGE and electroblotted to nitro-cellulose membranes. Proteins were detected by incubating the membrane with anti-Collagen I antibody at a concentration of 0.2-2 ug/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 peroxidase-conjugated secondary antibody at 1 ug/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.