兔抗鑰孔戚血藍素IgGRabbit Anti KLH IgG ELISA 說明書

兔抗鑰孔戚血藍素IgG檢測試劑盒

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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