Histone Cluster 1, H4A (HIST1H4A) BioAssay(TM) ELISA Kit (Human, Rat, Mouse)

Product Overview

• Article Name: Histone Cluster 1, H4A (HIST1H4A) BioAssay(TM) ELISA Kit (Human, Rat, Mouse)
• Biozol Catalog Number: USB-227383
• Supplier Catalog Number: 227383
• Alternative Catalog Number: USB-227383-96
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

The Histone Cluster 1, H4A (HIST1H4A) ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of HIST1H4A in serum, plasma, tissue homogenates, cell lysates and other biological fluids. This kit is suitable for use with human, mouse and rat samples. Detection Range: 0.156-10ng/ml Sensitivity: <0.052ng/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 HIST1H4A. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to HIST1H4A. 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 HIST1H4A, 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 HIST1H4A in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *227383A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *227383B: Standard, 2x1vial 227383C: Standard Diluent, 1x20ml *227383D: Detection Reagent A, 1x120ul *227383E: Detection Reagent B, 1x120ul 227383F: Assay Diluent A, 1x12ml 227383G: Assay Diluent B, 1x12ml 227383H: TMB Substrate, 1x9ml 227383K: Stop Solution, 1x6ml 227383L: Wash Buffer, 30X, 1x20ml Storage and Stability: Store *227383A, *227383B, *227383D and *227383E at -20C. Store all the other components at 4C. 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. Microplate reader with 450 10nm filter. 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 Sample Preparation and Storage: Serum: Use a serum separator and allow samples to clot for two hours at room temperature or overnight at 4C before centrifugation for 20 minutes at approximately 1000xg. Assay freshly prepared serum immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Plasma: Collect plasma using EDTA or heparin as anticoagulant. Centrifuge samples for 15 minutes at 1000xg at 2-8C within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, tissues were rinsed thoroughly in ice-cold PBS (0.02M pH 7.0-7.2) to remove excess blood and weighed before homogenization. The tissues were minced to small pieces and homogenized in 5-10ml of PBS with a glass homogenizer on ice (Micro Tissue Grinders works too). The resulting suspension was sonicated with an ultrasonic cell disrupter or subjected to two freeze-thaw cycles to further break the cell membranes. After that, the homogenates were centrifuged for 5 minutes at 2500*g. The supernatant was removed and assayed immediately or aliquoted and stored at -20C or lower. Cell Lysates: Cells must be lysed before assaying according to the following directions. 1. Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly). 2. Wash cells three times in cold PBS. 3. Resuspend cells in PBS (1*) and subject the cells to ultrasonication 4X (or freeze cells at