V-Set Domain Containing T-Cell Activation Inhibitor 1 (VTCN1) BioAssay™ ELISA Kit (Human)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Human V-Set Domain Containing T-Cell Activation Inhibitor 1 (VTCN1) ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of VTCN1 in human tissue homogenates, cell lysates and other biological fluids.

Detection Range:
0.156-10ng/ml

Sensitivity:
0.058ng/ml

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 VTCN1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to VTCN1. 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 VTCN1, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of VTCN1 in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*028807A: Microtiter Plate, 1x48 wells or 1x96 wells, Pre-coated; ready to use
*028807B: Standard, 2x1vial
028807C: Standard Diluent, 1x10ml or 1x20ml
*028807D: Detection Reagent A (green), 1x60ul or 1x120ul
*028807E: Detection Reagent B (red), 1x60ul or 1x120ul
028807F: Assay Diluent A, 1x6ml or 1x12ml
028807G: Assay Diluent B, 1x6ml or 1x12ml
028807H: TMB Substrate, 1x4.5ml or 1x9ml
*028807J: Positive Control, 0.77ng/ml, 1x1vial
028807K: Stop Solution, 1x3ml or 1x6ml
028807L: Wash Buffer, 30X, 1x10ml or 1x20ml

Precaution:
The Stop Solution (028807K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage and Stability:
Store *028807A, *028807B, *028807D, *028807E and *028807J at -20°C. After reconstitution of *028807J, 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.

Notes on 028807J: Positive Control:

Intended Use:
The positive control is from human sample known to contain the target protein. It will provide a positive result in the ELISA assay, which indicates that the kit is optimized and working properly.

Characteristics:
VTCN1 in human sample (lyophilized powder). Note: The human based material used in this product are screened for HIV, Hepatitis A, B, C, HTLV and Syphilis.

Expected Values:
0.77ng/ml

Usage:
Dissolve with 150ul standard diluent and let stand for 10 minutes. If powder is partly insoluble, shake gently until fully dissolved. Mix gently before use. In the experiment, add 100ul to an appropriate well of the plate and test its concentration along with standards and your specimens. Note: Upon receipt store Positive Control vial at -20°C. After reconstitution, store Positive Control vial at 4°C and use within 5 days.

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 90ul TMB Substrate. Incubate 15-25 minutes at 37°C.
8. Add 50ul Stop Solution. Read at 450nm immediately.