Rabbit polyclonal antibody raised against native Collagen Type I.
Native purified human and bovine placenta Collagen Type I.
This antibody reacts with most mammalian Type I collagens and has negligible cross-reactivity with Type II, III, IV, V or VI collagens. Non-specific cross-reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible.
Quality Control Testing:
Antibody Reactive Against Native Purified Protein.
ELISA (1:5000-1:50000) Western Blot (1:5000-1:50000) Immunohistochemistry (1:50-1:200) The optimal working dilution should be determined by the end user.
In 100 mM Na2B4O7, 75 mM NaCl, 5 mM EDTA, pH 8.0 (0.01% sodium azide)
Store at 4°C on dry atmosphere. After dilution, store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Western blot analysis is shown using Collagen Type I polyclonal antibody (Cat # PAB10190) to detect expression of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARgamma-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 Collagen Type I polyclonal 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). See Hazra et al. (2004) for additional details.
Immunohistochemical staining with Collagen Type I polyclonal antibody (Cat # PAB10190) was used at a 1 : 100 dilution to detect distal tubules in normal kidney tissue. Note the absence of staining of glomeruli. The antibody was reacted with antibody for 4 hours at room temperature followed by the addition of secondary antibody and substrate reaction. Tissue was formalin-fixed and paraffin embedded. No antigen retrieval was performed. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.