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Description
Human HDAC5 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected for analysis. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 conjugate are sequentially added to microwells pre-coated with a histone deacetylase 5 (HDAC5) 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 deacetylase 5 (HDAC5) 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 Deacetylase 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Histone deacetylase 5 is an enzyme encoded by the HDAC5 gene. Diseases associated with HDAC5 include Alzheimer's disease and isolated hyperhidrosis in individuals with normal sweat glands. Pathways involved include activated PKN1, which stimulates transcription of the AR (androgen receptor)-regulated genes KLK2 and KLK3, and the apelin signaling pathway. Gene ontology annotations associated with this gene include transcription factor binding and transcriptional core inhibitor activity. An important homologue of this gene is HDAC9. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 5
A great laptop stand
At first, I was skeptical of the price, especially when I opened up the box and what I was presented with was nothing but a thick chunk of shaped metal. However, now that I've been using this for a bit of time, I can attest to it's quality:
1. This thing is STURDY. One of my children is a curious (read: nosy) four year old who loves to inspect anything she can get her hands on. No amount of her typing, tugging, pulling or otherwise manipulation of the work laptop upon this stand has budged it in the slightest. Unless you are (an adult) intentionally trying to remove the laptop from the stand, your computer is very stable.
2. It's actually great for hiding wires. It has a hole in the back for tunneling wires towards/away from the computer, and this stand being the hunk of metal that it is, does a nice job obscuring such wires, which gives the stand a nice, clean look on a desk with a little bit of care. For some people, this matters a lot, which gives this stand an edge over the more architecturally sparse stands on Amazon.
3. The space between the "legs" of the stand and the "floor" where your computer rests is a great place to hide your phone while getting work done. I put mine on a charger and hide my phone there, which is really nice for limiting any tendencies to check my phone when I have a lull in my attention span. Since you can't see the phone when it rests there, but you can still hear it, I found myself pleasantly surprised that this stand could enable better productivity from my bad habits.
It's a good stand and it's worth the money, especially if you have dual monitors and/or you're trying to improve the ergonomics of your workspace.
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Reviewed in the United States on September 14, 2024
★★★★★ 5
Timeless and sturdy!
Such a huge fan of this company. We've bought multiple stands and loved them. I had the older more silver ones previously but got the black one for my newer Mac and it's awesome. The design is perfect. and it's helpful not just for the function of holding the stand, but it's helping keep the laptop cool by drawing heat away and also airflow/etc.
I had a guy in the office the other day come ask me if he could swap me for the black one because he loved it so much. lol.
Anyway, great stands. We've been buying from this company for almost 10 years. Solid. They look as new today as they did when we bought the first one.
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Reviewed in the United States on September 30, 2025
★★★★★ 5
Does what it needs to
-Solid
-One piece construction
-Great accentuation of my laptop
-Pleasant angle to still keyboard if you need to
-Rubberized feet keep it in place
-Reasonably priced
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Reviewed in the United States on October 8, 2025
★★★★★ 5
Works great in my setup
I got this for my work desk, and it does exactly what it's advertised to do: it holds my laptop (which is 16 inches). And it doesn't wobble and shake whenever I type on my external keyboard (I use a Unicomp Ultra Classic, which uses buckling springs, so I'm putting a decent amount of force into each keypress - if a stand is not stable, it will be immediately obvious to me), and I can even move my mouse underneath the stand without running into any problems. (Before, when my laptop was sitting directly on the desk, it was annoying how it made me move my keyboard around and inhibited my range of mouse movement, but now I can place my keyboard right where I want it and use my mouse as I please.)
The stand feels very sturdy and well-constructed, and it keeps my laptop at eye level. Height-adjust would be nice, but I have yet to find a good quality adjustable laptop stand, so I'll just use books to adjust it if the need arises. The box it comes in is also quite nice.
This thing freed up desk space where I needed it and is unlikely to ever fail (there are no moving parts), so I wholeheartedly recommend it to anyone looking for a laptop stand.
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Reviewed in the United States on July 8, 2023
★★★★★ 4
Beautiful design
Simple, functional and beautifully designed, I'm glad I bought it.
(not a five starts just because I thought it was higher, but again, my fault for not reviewing the specs first)
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Reviewed in the United States on October 29, 2024