Description
Mouse Anti-Porcine Type I Collagen IgG Antibody Assay Kit, TMB - Cat Number: 1013T From Chondrex.
Research Field: Arthritis, Immunology
Clonality: N/A
Cross-Reactivity:
Host Origin: N/A
Applications: N/A
Isotype: N/A
Detection Range: 16 units/ml-0.25 units/ml
Sample Type: Serum, Plasma
Concentration: N/A
Immunogen:
DESCRIPTION: Assay kit to quantify mouse anti-collagen antibodies
FORMAT: Pre-coated 96-well ELISA Plate with removeable strips
ASSAY TYPE: Indirect ELISA
ASSAY TIME: 4.25 hours
STANDARD RANGE: 16 units/ml to 0.25 units/ml
NUMBER OF SAMPLES: Up to 39 (duplicate) samples/standard plate (will vary for custom kits)
SAMPLE TYPES: Serum and Plasma
RECOMMENDED SAMPLE DILUTIONS: 1:1000 (at least)
CHROMOGEN: TMB (read at 450 nm)
STORAGE: -20°C
VALIDATION DATA: N/A
INTRODUCTION
This kit is designed to assay type I and type II collagen antibodies in mouse serum. Chondrex Inc.’s ELISA systems incorporate uniqueblocking agents to reduce non-specific reactions. These agents reduce the background levels by inhibiting the hydrophobic binding of immunoreactive serum components in sample specimens onto plastic surfaces. Chondrex, Inc. offers various species of type I and type II collagen-coated strips as shown below. This ELISA kit contains enough material to run two plates on two separate occasions. Heterologous type II collagen is widely used as an immunogen for the collagen-induced arthritis (CIA) model.
In CIA-susceptible mice, the serum antibody levels to the type II collagen used for immunization are very high. Furthermore, these antibodies cross-react to various species of type II collagen including autologous type II collagen, due to the conserved amino acid sequences of type II collagen. It is important to note that although type I collagen shares about 80% of its amino acid sequence with type II collagen, it is not capable of inducing autoimmune-mediated diseases. This indicates that epitope specificity of antibodies and T-cells are important for establishing autoimmunity and subsequent development of autoimmune diseases. Therefore, type I collagen might be useful as a control for studying the pathogenesis of autoimmune mediated arthritis and B and T-cell epitope specificity.
KIT COMPONENTS
PLATE COATING AND SETUP
NOTES BEFORE USING ASSAY
NOTE 1: It is recommended that the standard and samples be run in duplicate.
NOTE 2: Warm up all buffers to room temperature before use.
NOTE 3: Crystals may form in Wash Buffer, 20X when stored at cold temperatures. If crystals have formed, warm the wash buffer by placing the bottle in warm water until crystals are completely dissolved.
NOTE 4: Measure exact volume of buffers using a serological pipet, as extra buffer is provided.
NOTE 5: Cover the plate with plastic wrap or a plate sealer after each step to prevent evaporation from the outside wells of the plate.
NOTE 6: For partial reagent use, please see the assay protocol’s corresponding step for the appropriate dilution ratio. For example, if the protocol dilutes 50 µl of a stock solution in 10 ml of buffer for 12 strips, then for 6 strips, dilute 25 µl of the stock solution in 5 ml of buffer. Partially used stock reagents may be kept in their original vials and stored at -20⁰C for use in a future assay.
NOTE 7: This kit contains animal components from non-infectious animals and should be treated as potential biohazards in use and for disposal.