人抗鑰孔戚血藍素IgG Human Anti KLH IgG ELISA Kit 說明書

人抗鑰孔戚血藍素IgG檢測試劑盒

Human Anti Keyhole limpet hemocyanin (KLH) IgG ELISA Kit

cat#700-140-KLG

INTENDED USE

The Human Anti-KLH IgG ELISA Kit detects and quantifies KLH(keyhole limpet hemocyanin) -specific IgG in human serum orplasma of vaccinated, immunized and/or infected hosts. Thisimmunoassay is suitable for:o Determining immune status relative to non-immunecontrols;o Assessing efficacy of vaccines, including dosage,adjuvantcy, route of immunization and timing;o Qualifying and standardizing vaccine batches & protocols.o Measuring natural anti-KLH antibodies of innate immunity.This kit is for research use only (RUO) and not for diagnosis cureor prevention of the disease.

GENERAL INFORMATION

Keyhole Limpet Hemocyanin (KLH), an oxygen-transportingprotein of the marine gastropod Megathura crenulata, isrecognized as a potent immunoactivator, and therefore is widelyused in research and clinical studies. Present applications of KLHinclude: (a) use as a highly immunogenic antigen for assessmentof immune competence of an organism, and (b) frequent use as acarrier of low molecular mass peptides and haptens, such asoligosaccharides, gangliosides or (glyco)peptides, designed tofacilitate antibody production. In these cases, antibodies made tosmall peptides/haptens are generally raised by coupling to a largecarrier protein like KLH. Antibodies are produced to both KLH andthe peptide/hapten; because anti-KLH may give non-specificsignals in various immunoassays, removal by solid phaseimmunoaffinity chromatography is common. The ELISA is usefulfor determining levels of anti-KLH in sera and for monitoring antiKLH removal from purified samples after antibody production.The ADI anti-KLH ELISA is designed with high sensitivity fordiscriminating lower level antibodies, which may include naturalanti-KLH antibodies of innate immunity, with specially formulateddiluents to minimize interfering background signals.

PRINCIPLE OF THE TEST

The Human Anti-KLH IgG ELISA kit is based on the binding of humananti- KLH in samples to KLH antigen immobilized on the microwells,and anti- KLH IgG antibody is detected by anti-human IgG-specificantibody conjugated to HRP (horseradish peroxidase) enzyme. Aftera washing step, chromogenic substrate (TMB) is added and color isdeveloped by the enzymatic reaction of HRP on the substrate, which isdirectly proportional to the amount of anti-KLH IgG present in thesample. Stopping Solution is added to terminate the reaction, andabsorbance at 450nm is then measured using an ELISA microwellreader. The activity of human antibody in samples is determinedrelative to anti-KLH calibrators.PRODUCT

SPECIFICATIONS

SpecificityPharma grade keyhole limpet hemocyanin (KLH) is used to coatthe microwells; stabilizing postcoat contains BSA; thus, no otherantibody specificity is detectable in the assay. The anti-HumanIgG HRP conjugate specifically detects IgG, and will not react withIgM, IgA or IgE class antibodies.Assay SensitivityThe KLH-coated plated and the anti-Human IgG HRPconcentration are optimized to differentiate anti-KLH IgG frombackground (non-antibody) signal with human serum samplesdiluted 1:200.Calibrator ValuesThe Calibrators are composed of dilutions of IgG antibody toKLH. Values are assigned as arbitrary anti-KLH activity units(see Limits of the Assay).

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