兔抗钥孔戚血蓝素IgG检测试剂盒 Rabbit Anti Keyhole limpet hemocyanin (KLH) IgG ELISA Kit cat#700-110-KLR INTENDED USE
The Rabbit Anti-Keyhole Limpet Hemocyanin (KLH) ELISA Kit is an immunoassay suitable for quantifying or titering total antibody activity (IgG, IgA and IgM) specific for KLH in serum, plasma or other biological fluids. For research use only (RUO), not for diagnosis, cure or prevention of the disease. RESEARCH USE OF THE TEST Keyhole Limpet Hemocyanin (KLH), an oxygen-transporting protein of the marine gastropod Megathura crenulata, is recognized as a potent immunoactivator, and therefore is widely used in research and clinical studies. Present applications of KLH include: (a) use as a highly immunogenic antigen for assessment of immune competence of an organism, and (b) frequent use as a carrier of low molecular mass peptides and haptens, such as oligosaccharides, gangliosides or (glyco)peptides, designed to facilitate antibody production. In these cases, antibodies made to small peptides/haptens are generally raised by coupling to a large carrier protein like KLH. Antibodies are produced to both KLH and the peptide/hapten; because anti-KLH may give non-specific signals in various immunoassays, removal by solid phase immunoaffinity chromatography is common. The ELISA is useful for determining levels of anti-KLH in sera and for monitoring anti-KLH removal from purified samples after antibody production. The ADI anti-KLH ELISA is designed with high sensitivity for discriminating the lower level antibodies, with specially formulated diluents to minimize interfering background signals. PRINCIPLE OF THE TEST The Rabbit KLH ELISA kit is based on the binding of rabbit anti- KLH in samples to KLH immobilized on the microwells, and anti- KLH Ig antibody is detected by anti-rabbit Ig-specific antibody conjugated to HRP (horseradish peroxidase) enzyme. After a washing step, chromogenic substrate (TMB) is added and color is developed by the enzymatic reaction of HRP on the substrate, which is directly proportional to the amount of anti-KLH present in the sample. Stopping Solution is added to terminate the reaction, and absorbance at 450nm is then measured using an ELISA microwell reader. The amount of rabbit Ig in samples is calculated relative to rabbit anti-KLH reference calibrators. Specificity Purified KLH is used to coat the microwells; thus the assay is specific for antibodies directed to KLH. The anti-rabbit IgG (H+L) HRP conjugate reacts with rabbit IgG, IgA and IgM class antibodies that bind to KLH on the plate. IgE antibody would not be measured above background signals. Assay Sensitivity The KLH coating level and HRP conjugate concentration are optimized to differentiate anti-KLH Ig from background (non-antibody) signal with rabbit serum samples diluted 1:100. Calibrator Values The calibrators are dilutions of rabbit IgG antibody reactive to KLH. Values are assigned as arbitrary Anti-KLH activity units (see Limits of the Assay).
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