Purified collagens are prepared by controlled and limited pepsin digestion of either placental villi (types I, III, IV, V and Vl) or nasal cartilage (bovine type II), followed by selective salt precipitation to yield the various collagen species.

The purity of each collagen preparation is assessed by SDS-PAGE analysis.

All purified collagens are supplied as a 0.5mg/ml solution in 500mM acetic acid. The soluble collagen may be dialyzed into physiological buffers.

• Collagen standards
• Antigen for antibody production
• Coating material for cell culture studies
• Formation of collagen gels

Purified collagen may be stored at 2-8°C for one year without appreciable loss of binding activity. For long-term storage, collagen may be aliquoted and stored frozen at -20°C. Avoid repeated freezing and thawing.

Antibodies are isolated from high titer antiserum by affinity chromatography, following which the purified antibodies are extensively cross absorbed to remove unwanted specificities.

The specificity and activity of each affinity purified anti-collagen antibody is tested by ELISA to conform with characteristics of a standard reference reagent. Although the antibodies are specific for bovine and human collagen, the antibodies may cross react with collagen from other species.

Unlabeled antibodies. 0.2 mg affinity purified antibody in 0.5 mL (0.4 mg/mL) of 100 mM borate buffered saline, pH 8.2. No preservativesor amine-containing buffer salts added.

Biotin-labeled antibodies. 0.2 mg labeled antibody in 0.5 mL (0.4 mg/mL) of phosphate buffered saline, pH 7.4, containing 0.1% sodium azide as preservative.

Both unlabeled and biotin-labeled anti-collagen antibodies can be stored for several months at 2-8°C without appreciable loss of activity. For long term storage, the antibodies should be aliquoted and frozen at -20°C. Avoid repeatedfreezing and thawing.

• ELISA
• Immunoblotting
• Indirect immunofluorescence staining
• Immunohistochemistry
• Immunocytochemistry

NOTE: The anti-collagen antibodies are recommended for indirect immunofluorescence, immunocytochemical and immunohistochemical staining of frozen sections only. Most tissue fixation methods employed for most paraffin embedding procedures appear to denature the conformational collagen epitopes recognized by the antibodies

Unlabeled antibodies. Since affinity purified anticollagen antibodies can be used for a variety of applications, the optimal working dilution for a particular application should be determined by the investigator.

Biotin-labeled antibodies. Dilute 1:5,000 for ELISA, 1:500-1:1,000 for immunoblotting and 1:50-1:200 for immunocytochemistry or immunohistochemistry.