Mouse insulin autoantibodies,IAA ELISA Kit, 96T

Mouse insulin autoantibodies,IAA ELISA Kit, 96T

Purpose

This kit is used for the determination the existence of Mouse Insulin Auto-antibodies (IAA) in mouse serum, plasma, cell culture supernatants, and other biological fluids.

Principle of the assay

The kit assays mouse IAA in the samples by the method of purified IAA antigen pre-coated enzyme-labeled SPA ELISA. The solid-phase antigen is prepared by purified mouse IAA antigen which is pre-coated onto a microplate. After loading of samples, the IAA in the controls or samples bind with the coated antigen. Then the Staphylococcal protein A (SPA)-enzyme immunoglobulin conjugates has been added into wells. After washing to remove any unbound reagent, the tetramethylbenzidine (TMB) substrate is added. TMB substrate produces blue color when catalyzing by the HRP enzyme. After reaching the desired color intensity, the reaction is terminated by addition of an acidic Stop solution which changes the solution color from blue to yellow. Determining the absorbance (OD Value) at wave length 450 nm by the microwell reader, and the existence of IAA can be calculated according to OD from positive or negative controls.

Calculation

1.       Average the duplicate readings for each control and sample and subtract the average of blank well (1 x sample buffer only).

2.       Determine P/N value: P/N= OD_sample /OD_negative. If OD_negative is less than 0.10, please use 0.10 as the value. OD_positive or OD_negative is the OD₄₅₀ from positive or negative control.

3.       If P/N ≥ 2.1, the testing sample is IAA positive; if P/N < 2.1, the testing sample is IAA negative.

4.       The normal range of OD_negative should be less than 0.21, and OD_positive should be more than 0.40, otherwise the experiment may have some errors and maybe need repeating.

5.       If the OD_sample is more than 2.5 or less than 0.1, it is out of the optimal test range. Dilution or concentration of sample, or adjust incubating time may be necessary for accuracy.

Kit Performance

1.       Sample from healthy mouse should be negative.

2.       Specificity: simultaneous detection of recombinant or natural mouse IAA without cross-reaction with other cytokines.

3.       Repeatability: both inter-assay and intra-assay were less than 10 %.

*Lead time of this kit is 1-2 weeks. Shipped by FedEx air. 
*Storage: 2 - 8 C. Shelf Time: 6 months.
*Kit Experimental Time:< 2 hs

>>Trial pricing is available for first time use for certain customers, contact us to see if you are qualified.
>>>Need manual or more information of this products Email us or Call us (1-866-897-4664)

Important note: Products are intended for laboratory or manufacture use and are not to be used for diagnosis or any other purpose, which include but are not limited to, unauthorized commercial uses, ex vivo or in vivo therapeutic uses. Not for Human or Animal Consumption.