This ELISA kit is designed to assay type II collagen antibodies in rat sera. The microtiter strips provided in the kit have been coated with human type II collagen. To reduce non-specific binding, we have incorporated unique blocking agents which inhibit the hydrophobic binding of immunoreactive serum components in sample specimens onto plastic surfaces and thus reduce the background levels.
Collagen is an inert, rigid protein found predominantly in skin, ligaments, bones and teeth. Its most distinctive attribute, essential to a transmitter of mechanical force, is inelasticity. Its fundamental structural unit is tropo-collagen, a molecular rod about 2600Å in length and 15Å in diameter and 300,000 molecular weight. In tendons these macromolecules, grouped as collagen fibrils, run parallel to the axis. In skin the fibrils are interlaced and branched.
Heterologous type II collagen is widely used for inducing arthritis in susceptible animals. The serum antibodies in collagen-induced arthritic (CIA) mice are highly specific to the type II collagen used for immunization, however there is significant crossreactivity with other species of type II collagen including rat type II collagen.
Kit Components:1. Type II collagen-coated 8-well stripsa) Ten, 8 well strips coated with one species of type II collagen for assaying antibody levels in sample specimens. b) Four, 8 well strips for reference standards. Two strips are used per run allowing two partial plates to be run on two separate occasions
2. Standard antibody (1.2ml, 16units/ml)3. Solution A (Blocking buffer, 15ml)4. Solution B (Sample dilution buffer, 60ml): Use for diluting standards and sample specimens
5. Secondary antibody (two vials of lyophilized, peroxidase-conjugated goat anti-rat IgG): Dissolve one vial in 10ml of solution C (Secondary antibody dilution buffer, 20ml)6. Urea-H2O2 plus buffer tablet containing (two tablets): Dissolve one tablet in 20ml glass-distilled water
7. OPD tablet (two tablets): Dissolve one tablet in 20ml Urea-H2O2 buffer (see step 6 above), just before use
8. Solution D Stop solution: (2.5N Sulfuric acid, 7.5ml)9. Solution E 20X Wash buffer (50ml): dilute in 1 liter of distilled water.
