The Secretory Immunoglobulin A (sIgA) BioAssay™ ECL ELISA Kit (Bovine) is a sensitive chemiluminescent sandwich immunoassay for the in vitro quantitative determination of sIgA in serum, plasma, saliva and other biological fluids.
Detection Range:0.781-50ng/ml
Sensitivity:<0.22ng/ml
Precision:Intra-Assay: CV<10%Inter-Assay: CV<12%
Test Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to sIgA. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to sIgA. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The addition of substrates A and B mixture generates glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the sIgA level in the sample or standard.
Kit Components:*158098A: Microtiter Plate, 1x96 wells, Pre-coated with antibody; ready to use
*158098B: Standard, 2x1vial 158098C: Standard Diluent, 1x20ml*158098D: Detection Reagent A (green), 1x120ul *158098E: Detection Reagent B (red), 1x120ul 158098F: Assay Diluent A (2x), 1x6ml158098G: Assay Diluent B (2x), 1x6ml158098H: Substrate A, 1x10ml158098K: Substrate B, 1x2ml158098L: Wash Buffer (30x), 1x20ml
Storage and Stability:Store *158098A, *158098B, *158098D and *158098E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Materials Required But Not Supplied:1. Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0secs; read time 1.0 sec/well . 2. Precision single or multi-channel pipettes and disposable tips
3. Eppendorf Tubes for diluting samples
4. Deionized or distilled water
5. Absorbent paper for blotting the microtiter plate
6. Container for Wash Solution
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 100ul Substrate Solution. Incubate 5-10 minutes at 37°C
