| Chitinase-3-like protein 1, also called CHI3L1 or YKL-40 is a secreted glycoprotein that is approximately 40kD in size that in humans is encoded by the CHI3L1 gene. This gene is mapped to 1q32.1. Chitinases catalyze the hydrolysis of chitin, which is an abundant glycopolymer found in insect exoskeletons and fungal cell walls. The glycoside hydrolase 18 family of chitinases includes eight human family members. This gene encodes a glycoprotein member of the glycosyl hydrolase 18 family. The protein lacks chitinase activity and is secreted by activated macrophages, chondrocytes, neutrophils and synovial cells. The protein is thought to play a role in the process of inflammation and tissue remodeling. The Chitinase 3-like 1/YKL-40 BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity ELISA for quantitative detection of mouse Chitinase 3-like 1/YKL-40 in cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Detection Range: 62.5-4000pg/ml Sensitivity: <10 pg/ml Specificity: Recognizes natural and recombinant mouse Chitinase 3-like 1/YKL-40. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This The Chitinase 3-like 1/YKL-40 BioAssay(TM) ELISA Kit is a solid phase immunoassay specially designed to measure human Chitinase 3-like 1/YKL-40 with a 96-well strip plate that is pre-coated with antibody specific for Chitinase 3-like 1/YKL-40. The detection antibody is a biotinylated antibody specific for Chitinase 3-like 1/YKL-40. The capture antibody is a monoclonal antibody from rat and the detection antibody is a polyclonal antibody from goat. The kit contains recombinant mouse Chitinase 3-like 1/YKL-40 with immunogen: Expression system for standard: NS0, Immunogen sequence: Y22-A381. To measure mouse Chitinase 3-like 1/YKL-40, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS buffer and add TMB. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The density of the yellow product is linearly proportional to mouse Chitinase 3-like 1/YKL-40 in the sample. Read the density of the yellow product in each well using a plate reader, and benchmark the sample wells readings against the standard curve to determine the concentration of mouse Chitinase 3-like 1/YKL-40 in the sample. Kit Components: 143775A: Microtiter Strips, 1x96 wells *143775B: Chitinase 3-like 1/YKL-40, Standard, 2x10ng 143775C: Anti-mouse Chitinase 3-like 1/YKL-40 (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *Chitinase 3-like 1/YKL-40B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 minutes or at RT for 2 hours. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with Working Wash Buffer. 3. Add 100ul of ABC working solution and incubate the plate at 37C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with Working Wash Buffer. 4. Add 90ul of TMB color developing agent and incubate the plate at 37C in dark for 15-25 minutes or at RT for 30 minutes. 5. Add 100ul TMB Stop Solution and read. |
