小鼠抗胰島素IgG檢測Mouse Anti Insulin IgG ELISA Kit說明書

小鼠抗胰島素IgG檢測試劑盒

Mouse Anti Insulin IgG ELISA Kit

cat#3700-MIG

INTENDED USE

The Mouse Anti-Insulin IgG ELISA Kit is an immunoassay

suitable for quantifying or titering IgG antibody activity specific for

insulin in serum or plasma. Other biological fluids, including

tissue culture medium, may be validated for use.

GENERAL INFORMATION

Insulin-dependent diabetes mellitus, type 1 diabetes (T1D), is an

autoimmune disease in which the insulin-producing pancreatic

islets are destroyed by T lymphocytes. Studies in non-obese

diabetic (NOD) mice, an animal model for the human disease,

have shown that they develop spontaneous proliferative T cell

response to several  cell autoantigens, including insulin. It is

hypothesized that diabetes development in NOD mice is due to

the loss of tolerance to autoantigens, and a biased activation of

Th1 cells may enhance diabetes, whereas Th2 cells inhibit the

disease. In efforts to combat the autoreactive causes of T1D in

the NOD model, depletion of B cells early in development by

genetic or serological approaches and targeted elimination of

mature B cells, as well as interventions that target intracellular B

cell signaling, have been shown to provide protection against this

T cell-mediated disease.

This Mouse Anti-Insulin ELISA allows for detection and

quantitation of even very low levels of anti-insulin IgG by

providing a high discrimination between background and true

anti-insulin signals. The anti-IgG HRP conjugate also produces

high signals with the IgG2c subclass of IgG in NOD mice, which

do not produce the IgG2a subclass.

PRINCIPLE OF THE TEST

The Mouse Anti-Insulin IgG ELISA kit is based on the binding of

mouse anti- insulin IgG in samples to insulin immobilized on the

microwells, and anti-Insulin IgG is detected by anti-mouse IgG

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-Insulin IgG 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

mouse IgG antibody in samples is calculated relative to mouse

anti-Insulin calibrators.

PRODUCT SPECIFICATIONS

Specificity

Purified human and bovine insulin are used to coat the

microwells; thus the assay is specific for antibodies directed to

insulin. The anti-Mouse IgG HRP conjugate reacts specifically

with mouse IgG; IgA, IgM and IgE class antibodies would not be

measured above background signals.

Assay Sensitivity

The insulin coating level, HRP conjugate concentration and Low

NSB Sample Diluent are optimized to differentiate anti- insulin

IgG from background (non-antibody) signal with mouse serum

samples diluted 1:50.

Calibrator Values

The calibrators are dilutions of mouse antibody reactive to

Insulin. Values are assigned as arbitrary anti-Insulin activity units

(see Limits of the Assay).

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