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Description
Human S100A5 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 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 utilizes 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 S100 Calcium Binding Protein A5 (S100A5) 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 S100 Calcium Binding Protein A5 (S100A5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human S100 Calcium Binding Protein A5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | S100 calcium-binding protein A5 (S100A5) is a protein encoded by the S100A5 gene in humans. The protein encoded by this gene is a member of the S100 family and contains two EF-hand calcium-binding motifs. S100 proteins are localized in the cytoplasm and/or nucleus of various cell types and participate in the regulation of cellular processes, such as cell cycle progression and differentiation. The S100 gene group includes at least 13 members, which form a cluster on chromosome 1q21. This protein has a Ca2+ affinity 20- to 100-fold higher than other S100 proteins studied under the same conditions. This protein also binds Zn2+ and Cu2+, with Cu2+ strongly affecting Ca2+ binding. Expression of this protein in the adult brain is very restricted. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
Great addition to our coffee station.
Size: EK6002
Love it. It's accuracy might be a little fuzzy about measuring the last tenth of a gram, probably due to rounding. But it showed a difference when a single coffee bean was added. Easy to read. The removable pad makes it easier to clean. It fits right on my little coffee station. To be honest, I didn't read the instructions. It's fairly intuitive if you know how they are supposed to work. I like it.
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Reviewed in the United States on November 26, 2025
★★★★★ 1
Broken, after 1 week of use
Size: EK6002
Sorry but, no. Stopped working after less than 1 week of use. They pre-install cheap batteries. I used it for a few days but then It wouldn’t turn on so I switched them out with fresh ones. Now the start/stop button isn’t working. Discovered during an extraction…arg!! Don’t mess with me while performing my craft! lol. Build quality is poor. Don’t over handle it, yeah, that’s how fragile it appears to be. I like that it is light weight but so light, I question if the durability and weight factor are compromising the output accuracy. Doesn’t feel reliable at all. I also question if this is waterproof, again, having everything to do with build quality. This is not my first scale but it is for coffee use so I’m on the hunt for a better one. This was cheap in price, yes. But as the old saying goes, you get what you pay for. One good thing that came out of this is, while it was working, I realized how much this was really needed for me to make exceptional and accurate espresso shots, so time to find a better one. I wanted this to work, I really did.
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Reviewed in the United States on April 3, 2026
★★★★★ 5
4.95 stars
Color: Black
TLDR: Buy it. It works well, it has impressive and useful features, it looks great, it’s thin/low-profile, and it’s a great price.
Detailed review: I’m very happy with this scale, and it has certainly upped my espresso game by leaps and bounds. I’m able to weigh my beans very precisely down to a tenth of a gram (roughly the weight of a single bean), so my dosing is on point. Also, it fits beneath the portafilter with room to spare for my cup, so I can weigh my shots and accurately measure my input/output ratio. And automatic mode is magic! When the espresso hits the cup, the timer starts running, so I can measure flow rate along with everything else! Overall, super easy to use and very necessary for the at-home barista.
One caveat: It acted weird on the 2nd or 3rd day, and I thought I was going to have to return it. The weight would slowly creep up even though I wasn’t adding any coffee. I dusted it off, made sure it was dry, set it on a more stable surface… I still don’t know what the matter was, but it stopped freaking out and it hasn’t really given me any trouble since. Also, I should mention that the automatic function doesn’t always work 100% perfectly; it works, and usually works quite well, but depending on the flow rate, you might get some drops dribbling into the cup and the timer not detect it right away. (I think the timer must be activated according to some sort of algorithm that looks for a certain increase in a certain amount of time, so a slower initial drip rate might not trigger it at first.) Also weirdly, it seems like about half of the time when the weight stops increasing (i.e., espresso stops flowing) , the timer pauses (as I think it’s supposed to do), but other times, the timer keeps running until you manually stop it. I don’t know… It’s a weird glitch, I guess, but it doesn’t really make any difference. The weight is accurate, and that’s the primary thing to keep an eye on.
So, is it perfect? No, I guess not. But, it works extremely well and functions perfectly for my purposes. In fact, it’s a lot nicer and fancier than I expected – plus, it’s more affordable and more streamlined than its obvious rival the Acacia.
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Reviewed in the United States on June 1, 2024
★★★★★ 5
Excellent and super accurate
Color: Black, Color: Black
I had a precision oxo which was more expensive than this one but not great though, it would register .4 grams suddenly and not sensitive to the 1-2 bean increments. I bought this one for espresso specifically and man it is fantastic so accurate, super responsive, well built, size is great if you want to weigh your shot under your portafilter. I have a fellow series 1 and it’s amazing working with it. Overall highly recommend and for the price it’s amazing great value! Not sure why would I buy a bookoo for $130 or a fellow scale for $200. This one works great for $40
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Just a tiny, accurate scale
Color: Black
Works flawlessly after I calibrated it compared to my other. Out of the box it was (very) off. Now it's within .1 of a gram. It's far more responsive than I expected. It's perfect for espresso and it's tiny so it fits on the smallest drip trays.
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Reviewed in the United States on May 8, 2026