Histone H2A ELISA Kit

Catalog No. ABIN6956582

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. Due to the 100% homology of the sequence among different species, the kit can be used to 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)

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

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:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

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.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target 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.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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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