Signal Transducer And Activator Of Transcription 3 (STAT3) BioAssay™ ELISA Kit (Human, Mouse, Rat)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Signal Transducer And Activator Of Transcription 3 (STAT3) ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of STAT3 in human, mouse and rat tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.

Detection Range:
1.56-100ng/ml

Sensitivity:
<0.61ng/ml

Specificity:
This assay has high sensitivity and excellent specificity for detection of STAT3. 100% cross-reactivity was observed among human, mouse and rat.

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 STAT3. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to STAT3. 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 STAT3, 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 STAT3 in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*366550A: Microtiter Plate, 1x96 wells, Pre-coated; ready to use.
*366550B: Standard, 2x1vial
366550C: Standard Diluent, 1x20ml
*366550D: Detection Reagent A, 1x120ul
*366550E: Detection Reagent B, 1x120ul
366550F: Assay Diluent A, 1x12ml
366550G: Assay Diluent B, 1x12ml
366550H: TMB Substrate, 1x9ml
366550K: Stop Solution, 1x6ml
366550L: Wash Buffer, 30X, 1x20ml

Storage and Stability:
Store *366550A, *366550B, *366550D and *366550E 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.

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.