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Description
Human S100A2 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 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 S100 Calcium Binding Protein A2 (S100A2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of S100 Calcium Binding Protein A2 (S100A2) 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 A2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | S100 calcium-binding protein A2 (S100A2) is a protein encoded by the S100A2 gene in humans, which is located on chromosome 1q21 along with other S100 proteins. S100A2 is also known as CaN19 or S100L. Under normal conditions, it is highly expressed in the human lung, prostate, kidney, hair follicles, salivary glands, and mammary glands. S100A2 is primarily localized in the cell nucleus, which is less common than other S100 proteins. It can also be found in the cytoplasm, where its distribution is quite diffuse. Its cytoplasmic presence is likely dependent on cellular calcium levels. In the extracellular environment, it can be found as a homodimer both in vitro and in vivo, but it also exists in monomeric, aggregated, and multimeric forms. In its multimeric form, it functions as a ligand for the RAGE receptor. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.2 ★★★★★
Based on 219 reviews
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Product Reviews
★★★★★ 5
Gorgeous 😍
Size: 2' x 6' (Natural), Color: Champagne, Size: 2' x 6' (Natural), Color: Champagne
Absolutely beautiful!!! It is thick and luxurious, super soft and cozy. It’s really high quality and it shows. You can tell it’s natural without the burn test even from its cute little curls. All brushed (your pet brush is perfect 😉) out it looks stunning and fluffy. I got the champagne color and I love it! I don’t see any seams on the double pelt and it is substantial, not at all like it will disintegrate in a few years, but rather will become a much loved family heirloom. Well worth the investment! It is absolutely heavenly to perform yoga on by the fire 😌I love taking my slippers off and feeling the soft fur. I don’t want to get off it lol. I understand all those other customer’s kids and cats 🤣😂🤣
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Reviewed in the United States on November 26, 2023
★★★★★ 5
This is the one you want...
Size: 2' x 6' (Natural), Color: Natural (Yellow Undertones), Size: 2' x 6' (Natural), Color: Natural (Yellow Undertones)
I bought a ‘Genuine Sheepskin Rug Double Pelt Natural White Fur 2x6’ from Amazon in 2011 for $112.49, and recently decided to recommission it in my new living room. It looked so nice that I decided to add a second rug, and was pleased to find what Amazon claimed to be the same rug still available for only $83.78. Now I’m not one of those silly folks who expects the color and/or size of a natural product like a sheepskin rug to be perfectly predictable, and my living room is large enough that I didn’t need the two rugs to match perfectly, but when the new rug arrived, I couldn’t have been more disappointed. It was about 40% smaller than my older rug, was way, way off from anything that could be called white, and looked cheap, old, and tattered. I tried to comb it out a bit to see if I could make it look presentable, but it was a lost cause.
Amazon to the rescue. I returned the rug to Amazon for a full refund the next morning, and the transaction couldn’t have been simpler. And I immediately ordered the ‘Genuine Sheepskin Rug Double Pelt Natural White Fur 2x6’ shown on this page from Super Area Rugs for $119.00. What a difference! These rugs are large, plush, and super soft. The quality of these rugs is obvious, much, much nicer that either my old sheepskin rug or the ‘bargain’ rug I returned to Amazon. I was so impressed with the quality of this rug that I immediately ordered a second, which arrived this morning (it matches the other rug perfectly). My older sheepskin has been reassigned to another room because, well, it’s not in the same league as my two new rugs, which look terrific together.
As is so often the case, you get what you pay for. So if you’re looking for a top quality sheepskin rug, this is the one you want.
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Reviewed in the United States on December 23, 2015
★★★★★ 4
Very nice fleeces
This is a clean, soft, well groomed fleece intended for use on a 6 foot long sofa, so bought 2. They moved too much as a result of shifting bodies on the leather couch, so will get a longer version, sewn bottom to bottom, and used each of these fleeces on both armrests, instead.
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Reviewed in the United States on August 15, 2025
★★★★★ 5
Excellent Product!!! Will purchase again!
Size: 2' x 3' (Natural), Color: Wolf Tipped
Instead of ordering a cover for reclining loveseats, I chose to drape one of these over the headrest on either end of our loveseat. Not only do they protect the leather, they also help to elevate our decor. The rugs were received in a very timely manner. They are super soft, & warming as we move into the winter months. We've had family & friends comment on how great they look. 2 of them have already ordered, & added them to their sofa/loveseat/recliner decor. Have an issue of them slipping, add some velcro in between the crook of the cushion & the back of the furniture piece. It doesn't show & will help to keep the rug in place; if your busy body.!
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Reviewed in the United States on December 10, 2025
★★★★★ 5
Perfect for sensory sensitive kids and for the pure beauty of it
Size: 2' x 6' (Natural), Color: Natural (Yellow Undertones), Size: 2' x 6' (Natural), Color: Natural (Yellow Undertones)
After speaking over the phone with the seller, I was confident in this purchase for my SPD son. They do not use harsh chemicals or dyes in the process. It is from New Zealand and the only use in Vietnam is for shipping. My son hasn't been diagnosed yet for Autism but he is in the process. He is super sensitive to all things sensory and Amazon feedback isn't the place to give specific details on his condition but I will write that when he fist touched it- which was willingly- he smiled and asked to sleep on it. When he laid down on it and I covered him- he was calm and smiling. In the pic I provided he is calm and holding his door (his obsession is doors and windows). He even set his door down to hold the fur. For other mommies and papas with spd/autistic kids you know that willingly touching a new fabric and setting down an obsessive item is huge and super encouraging!
I am so grateful to have such a quality item that is affordable for my special needs son. His older sister received one as well as is now pretending to be a sheep. Fun for everyone!
The ONLY negative is that the brand/info sticker on the pelt was a pain to get off and pulled some of the felt - not damaging it but blemishing the unity -but if the skin is down it will not be noticeable.
A super fun detail is that all of the fur is combed out but there were a few strands that were wavy and had a slight curl- I LOVED that.
There was no odor at all which is great! I expected an animal smell which is better than chlorine or dioxin which is what competitors use in their process.
There is no noticeable seam in the middle on the top where the two pelts are gathered.
I am soooo satisfied!
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Reviewed in the United States on February 13, 2016