Histone H2A ELISA Kit

Catalog No. ABIN1874257

Product Details Histone H2A ELISA Kit

Target: Histone H2A

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.31 ng/mL - 20 ng/mL

Minimum Detection Limit: 0.31 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of H2A in tissue homogenates, cell lysates and other biological fluids. Due - the 100% homology of the sequence among different species, the kit can be used - detect human, mouse and rat samples.

Sample Type: Cell Lysate, Tissue Homogenate

Analytical Method: Quantitative

Specificity:

This assay has high sensitivity and excellent specificity for detection of Histone H2A (H2A).
No significant cross-reactivity or interference between Histone H2A (H2A) and analogues was observed.

Cross-Reactivity (Details): No significant cross-reactivity or interference between Histone H2A (H2A) and analogues was observed.

Sensitivity: 0.117 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Application Notes:

• Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
• The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
• Kits from different batches may be a little different in detection range, sensitivity and color developing time.
• Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
• Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
• There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
• Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
• Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
• Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
• Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Histone H2A (H2A). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Histone H2A (H2A). 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 Histone H2A (H2A), 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 Histone H2A (H2A) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 20 ng/mL. Prepare 7 tubes containing 0.5 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Histone H2A (H2A) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Histone H2A (H2A) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

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

Handling Advice: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Storage: 4 °C

Storage Comment:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Expiry Date: 6 months

Target Details for Histone H2A

Target: Histone H2A

Alternative Name: Histone H2A (H2A) (Histone H2A Products)

Synonyms: 5499 ELISA Kit, CG5499 ELISA Kit, Dmel\\CG5499 ELISA Kit, H2A ELISA Kit, H2A.F/Z ELISA Kit, H2A.X ELISA Kit, H2A.Z ELISA Kit, H2AV ELISA Kit, H2AV_DROM ELISA Kit, H2AX ELISA Kit, H2AZ ELISA Kit, H2Av ELISA Kit, H2AvD ELISA Kit, H2a.V ELISA Kit, H2av ELISA Kit, H2avD ELISA Kit, HIS ELISA Kit, HIS2AV ELISA Kit, HIS2AVD ELISA Kit, His ELISA Kit, His2 ELISA Kit, His2AV ELISA Kit, His2AvD ELISA Kit, HisH2Av ELISA Kit, Hist ELISA Kit, Hist2av ELISA Kit, gamma-H2Av ELISA Kit, gamma-HIS2AV ELISA Kit, gamma-His2Av ELISA Kit, gammaH2Av ELISA Kit, h2AvD ELISA Kit, his ELISA Kit, l(3)05146 ELISA Kit, l(3)810 ELISA Kit, l(3)97Dd ELISA Kit, l(3)L1602 ELISA Kit, DDBDRAFT_0205974 ELISA Kit, DDBDRAFT_0216271 ELISA Kit, DDB_0205974 ELISA Kit, DDB_0216271 ELISA Kit, RGD1564767 ELISA Kit, H2a-221 ELISA Kit, H2A-III ELISA Kit, HIST1H2AA ELISA Kit, Histone H2A variant ELISA Kit, putative histone H2A ELISA Kit, histone H2A ELISA Kit, histone cluster 1 H2A family member F ELISA Kit, histone H2A-like ELISA Kit, histone cluster 1, H2af ELISA Kit, histone2A1 ELISA Kit, histone cluster 1, H2A, III ELISA Kit, histone cluster 1, H2ac ELISA Kit, His2Av ELISA Kit, LMJF_17_0280 ELISA Kit, H2AX ELISA Kit, Hist1h2af ELISA Kit, LOC100425378 ELISA Kit, his2a1 ELISA Kit, HIST1H2A3 ELISA Kit, HIST1H2AC ELISA Kit, h2a-A ELISA Kit