Sample
Solubilized collagen and cell culture media
Intended Use
The Type II Collagen Detection ELISA Kit is designed to quantify the amount of solubilized native type II collagen from various species such as human, monkey, porcine, bovine, rat, mouse, rabbit, equine, dog, and chick in cell culture samples or tissue specimens by ELISA.
Contents of Kit
1. Type lI Collagen Standard, 1 vial, 100μl(100 μg/ml), -20°C
2. Capture Antibody, 1 vial, 100 μl(5 mg/ml), -20°C
3. Detection Antibody, 1 vial, Lyophilized, -20°C
4. Solution A-Coating Buffer, 1 bottle, 10 ml, -20°C
5. Solution B-Sample/Standard Dilution Buffer, 1 bottle, 50 ml, -20°C
6. Solution C-Detection Antibody Dilution Buffer, 1 bottle, 10 ml, -20°C
7. Solution D-Streptavidin Peroxidase Dilution Buffer, 1 bottle, 20 ml, -20°C
8. Streptavidin Peroxidase, 2 vials, 50μl, -20°C
9. OPD, 2 vials, Lyophilized, -20°C
10. Chromogen Dilution Buffer, 1 bottle, 20 ml, -20°C
11. Stop Solution-2N Sulfuric Acid, 1 bottle, 10 ml, -20°C
12. Wash Buffer 20×, 1 bottle, 50 ml, -20°C
13. ELISA Plate 1each 96-well (8-well strips × 12), -20°C
Storage
-20°C for 12 months
General Description
Type II collagen is unique among the collagen family, and its tissue distribution is limited to avascular tissues such as cartilage and the vitreous body of the eye. As type II collagen can induce arthritis in experimental animals, autoimmunity to type II collagen is suspected in the pathogenesis of certain autoimmune diseases in humans such as rheumatoid arthritis (RA), eye diseases associated with RA, and relapsing polychondritis, which affects specific tissues containing type II collagen. The Type II Collagen Detection ELISA Kit is designed to quantify the amount of solubilized native type II collagen from various species such as human, monkey, porcine, bovine, rat, mouse, rabbit, equine, dog, and chick in cell culture samples or tissue specimens by ELISA.
Standard Curve
1. Average the duplicate OD values for the blank (B), standards, and test samples.
2. Subtract the averaged blank OD values from the averaged OD values of the standards and test samples.
3. Plot the OD values of standards against the concentration of standards (ng/ml). Using a log/log plot will linearize the data. Figure 1 shows a representative experiment where the standard range is 3.1 - 200 ng/ml and the 1-step protocol and 2-step protocol are compared.
4. The ng/ml of collagen in test samples can be calculated using regression analysis. Multiply it by the sample dilution factor to obtain the collagen concentration (ng/ml) in the original sample specimens.
Citations
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COA
Certificate of Analysis
