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Description
Human H4 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with Histone H4 (H4) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Histone H4 (H4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Histone H4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Histone H4 is one of the five major histone proteins involved in chromatin structure in eukaryotic cells. H4 has a major globular domain and a long N-terminal tail, contributing to the "beads-on-a-string" organization of the ribosome. Histones are highly post-translationally modified. Covalent modifications include acetylation and methylation of the N-terminal tail. These modifications may alter gene expression on DNA associated with its parent histone octamer. Histone H4 is a crucial protein in chromatin structure and function, and its sequence variants and variable modification states are thought to play a role in both dynamic and long-term gene regulation. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
Perfect Prom Suit
Size: Small, Color: Dark Black, Size: Small, Color: Dark Black
This suit was perfect for prom. The fit was clean, the material looked high quality, and the texture gave it a unique look. Definitely worth it.
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Reviewed in the United States on April 23, 2026
★★★★★ 5
Award winning tuxedo!
Size: Large, Color: Baby Pink, Size: Large, Color: Baby Pink
Love this tuxedo suit! I’ve worn it a few times - to two different weddings and an Academy Awards viewing party, where I even won for best dressed. I get so many comments on it. The jacket and vest fit perfectly, however the pants are long and I had to hem them, but “stitch witchery” tape worked fine. The pants are also a little “stiff” - not a smooth fabric. But for occasional wear, it’s fine! The bow tie was sold separately.
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Reviewed in the United States on May 23, 2026
★★★★★ 5
Slim fit
Size: Large, Color: Brown
I recently purchased the brown slim-fit suit and am very impressed! For reference, I’m 6’2” and 222 lbs, and while I usually wear a size large, I went with an XL based on the fit advice and it was the perfect choice. This suit does run a bit small, so ordering a size up is definitely the way to go.
The slim fit is flattering and modern without being restrictive, and the rich brown color adds a unique and stylish touch to any occasion. The material feels high-quality, and the overall craftsmanship is excellent. If you're looking for a sharp, well-fitting suit, this one is a fantastic option—just be mindful of the sizing!
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Reviewed in the United States on January 14, 2025
★★★★★ 5
Good Decision !
Size: Large, Color: Dark Black, Size: Large, Color: Dark Black
Upon receiving the suit we were definitely skeptical because certain materials don't match the pictures. But Baby when we say this one came and showed up. It was great material , not cheap feeling if you get what Im saying. The pants, jacket and vest all came as it said it would. Nothing was missing! It came pressed and very wrinkle free so it was taking care of prior to shipping out. We got a Large for my son because of course he wanted a slimmer fit. But it did work out. He had enough room to comfortably be able to move. And for it truly was a steal for how much it costed. This literally felt like it was suppose to cost a few hundred , but it was so affordable.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
"Showstopper Suit That Turned Heads All Night!”
Size: Medium, Color: Violet Purple, Size: Medium, Color: Violet Purple
I absolutely love this suit. We bought it specifically to match my son's girlfriend's prom dress, and it could not have been a more perfect choice. After she picked out her gown, I spent the entire Sunday searching for a suit that would complement it, and when I saw this one, I knew it was the one. The colors, the pattern, the style—it all came together perfectly.
When prom day came, my son looked so confident, handsome, and proud. He and his date looked stunning together, like they coordinated their outfits down to the smallest detail. You could tell they were a couple just by the way they complemented each other. He received nonstop compliments, not only at the prom send-off but even later that night when we went to Texas Roadhouse. People were turning their heads and praising the look. It was truly a moment.
This was hands down one of the best purchases I have ever made on Amazon. I do not usually write reviews, but this suit deserves the recognition. It was sleek, bold, well-fitted, and absolutely top tier. If you're considering this suit, do it. It is a total showstopper.
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Reviewed in the United States on May 4, 2025