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

人抗鑰孔戚血藍素IgM檢測試劑盒

Human Anti  Keyhole limpet hemocyanin (KLH) IgM ELISA Kit

cat#700-145-KLM

INTENDED USE

The Human Anti-KLH IgM ELISA Kit detects and quantifies KLH (keyhole limpet hemocyanin) -specific IgM in human serum or plasma of vaccinated, immunized and/or infected hosts. This immunoassay is suitable for:

1)Determining immune status relative to non-immune controls;

2)Assessing efficacy of vaccines, including dosage, adjuvantcy, route of immunization and timing;

3)Qualifying and standardizing vaccine batches & protocols.

4)Measuring natural anti-KLH antibodies of innate immunity.

GENERAL INFORMATION

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 acarrier 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 lower level antibodies, which may include natural anti-KLH antibodies of innate immunity, with specially formulated diluents to minimize interfering background signals.

PRINCIPLE OF THE TEST

The Human Anti-KLH IgM ELISA kit is based on the binding of human anti-KLH in samples to KLH antigen immobilized on the microwells, and anti- KLH IgM antibody is detected by anti-human IgM-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 IgM 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 activity of human antibody in samples is determined relative to anti-KLH calibrators.

PRODUCT SPECIFICATIONS

Pharma grade keyhole limpet hemocyanin (KLH) is used to coat the microwells; stabilizing postcoat contains BSA; thus, no other antibody specificity is detectable in the assay. The anti-Human IgM HRP conjugate specifically detects IgM, and will not react with IgG, IgA or IgE class antibodies.

Assay Sensitivity

The KLH-coated plated, anti-Human IgM HRP concentration, and Low NSB Sample Diluent are optimized to differentiate anti-KLH IgM from background (non-antibody) signal with human serum samples diluted 1:200.

Calibrator Values

The Calibrators are composed of dilutions of antibody to KLH. Values are assigned as arbitrary anti-KLH activity units (see Limits of the Assay).

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