Histone H2B (H2B) BioAssay™ ELISA Kit (Human, Mouse, Rat)

Blue Ice

38220000

The Histone H2B (H2B) ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of H2B in tissue homogenages, cell lysates and other biological fluids. Due to 100% sequence homology among different species, this kit can be used for the detection of H2B in human, mouse and rat samples.

Detection Range:
3.12-200ng/ml

Sensitivity:
1.28ng/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 H2B. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to H2B. 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 H2B, 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 H2B in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*025705A: Microtiter Plate, 1x96 wells, Pre-coated; ready to use
*025705B: Standard, 2x1vial
025705C: Standard Diluent, 1x20ml
*025705D: Detection Reagent A, 1x120ul
*025705E: Detection Reagent B, 1x120ul
025705F: Assay Diluent A, 1x12ml
025705G: Assay Diluent B, 1x12ml
025705H: TMB Substrate, 1x9ml
025705K: Stop Solution, 1x6ml
025705L: Wash Buffer, 30x, 1x20ml

Storage and Stability:
Store *025705A, *025705B, *025705D and *025705E 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.