Cytochrome C Oxidase Subunit IV Isoform 2 (COX4I2) BioAssay™ ELISA Kit (Human)

The BioAssay™kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of COX4I2 in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.

0.156-10ng/ml

0.059ng/ml

This assay has high sensitivity and excellent specificity for detection of Cytochrome C Oxidase Subunit IV Isoform 2 (COX4I2). No significant cross-reactivity or interference between Cytochrome C Oxidase Subunit IV Isoform 2 (COX4I2) and analogues was observed.

Intra-Assay %CV: <10%
Inter-Assay %CV: <12%

The microplate provided in this kit has been pre-coated with an antibody specific to COX4I2. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to COX4I2. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain COX4I2, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of COX4I2 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Pre-coated, ready to use 96-well strip plate
Standard, 2x vials
Detection Reagent A, 1x120ul
Detection Reagent B, 1x120ul
TMB Substrate, 1x9ml
Wash Buffer (30X): 1x20ml
Standard Diluent, 1x20ml
Assay Diluent A, 1x12ml
Assay Diluent B, 1x12ml
Stop Solution, 1x6ml

Store Microtiter Strips, Standard, Detection Reagents and Positive Control at -20°C. After reconstitution of Positive Control, store at 4°C and use within 5 days. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. 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.

1. Prepare all reagents, samples and standards.
2. Add 100ul standard or sample to each well. Incubate 1 hour 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 90ul Substrate Solution. Incubate 10-20 minutes at 37°C.
8. Add 50ul Stop Solution. Read at 450nm immediately.

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.