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Description
Human MMP-8 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 125 ng/mL). Then dilute to the following concentrations: 125 ng/mL, 62.5 ng/mL, 31.25 ng/mL, 15.625 ng/mL, 7.8125 ng/mL, 3.90625 ng/mL, 1.953125 ng/mL, 0.01 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 125ng/mL standard working solution into the first EP tube and mix thoroughly to make a 62.5ng/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 a matrix metalloproteinase 8 (MMP-8) 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 matrix metalloproteinase 8 (MMP-8) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Matrix Metalloproteinase 8 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Matrix metalloproteinase-8 (MMP-8), also known as neutrophil collagenase, is a collagen-cleaving enzyme found in the connective tissues of most mammals. This gene is part of the MMP gene group, located on chromosome 11q22.3. Most MMPs are secreted as inactive proproteins and become activated when cleaved by extracellular proteases. However, the enzyme encoded by this gene is stored in secondary granules within neutrophils and is activated by autolytic cleavage. Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of the extracellular matrix during normal physiological processes such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes such as arthritis and metastasis. The primary function of MMP-8 is the degradation of types I, II, and III collagen. | |||||||||||||||||||||||||||||||||
| 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 | 0.19-12.5 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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Pretty Tunic Top
Color: Willowfloral, Size: X-Large
Very happy with this tunic top! Long enough for me to feel good about wearing it with leggings, but not SO long it feels like a dress. Really good V-neck, not just a little notch at the top that some brands call a v-neck. I ordered the teal floral XL and was very happy with the fit and drape. I'm a size 18W with a 42D bust and a chubby waistline. I often end up in XXL or 1X tops, but this fit well as an XL. Nice feel to the material and a bargain at the price!
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Reviewed in the United States on March 26, 2026
★★★★★ 5
Very flattering in my opinion
I'm 5'6 and no hips, so I really liked that the length is a little longer and flares a little at the hips giving my chunky but not curvy figure some illusion of a girlish figure. The pattern and colors were pretty & I'd say pretty close to the ad. Great blues and a coral/peach ... not sure my camera did it justice. I like the pleat at the back ... I feel that it minimizes the bra bulge that can happen at times. I stepped out after try on and my guy said it looked great (he's a truth teller with clothes) so that made me feel better because sometimes I see what I want to see at first & later see the flaws. I do see the case instructions ... getting used to hang to dry all things lately. Feels a little more nylon-ish but not bad. Its not super thin and light where I think it's a clingy thing. Really do like this shirt
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Reviewed in the United States on March 9, 2026
★★★★★ 4
Nice, but not great fabric quality
Color: Floral Black, Size: Large
The fit was great. Colors were as shown. The fabric shows signs of wear pretty much on the second wear so durability isn’t great. The fabric is soft and light weight so perfect for spring and fall. Not sure I’d buy again as I think it will look worn out quickly.
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Very Straightforward and Easy to Use
Reload type: One Time Reload
I have used this several times, and it has always been a smooth and convenient experience.
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It is especially useful for managing spending, setting a shopping budget, or keeping funds ready for future purchases. Instead of entering payment details every time, having balance available makes checkout faster and more convenient.
I also appreciate how quickly the funds are applied, allowing immediate use once the reload is completed.
This is one of those simple Amazon features that works exactly the way it should.
Overall, easy, efficient, and user-friendly. I have been very satisfied with it. Five stars.
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Reviewed in the United States on April 21, 2026
★★★★★ 5
“ incredibly convenient”
Reload type: One Time Reload
I recently used the Amazon.com Gift Card Balance Reload option, and I’m thoroughly satisfied with the process and the overall experience. Here’s my detailed review:
Convenience
The Gift Card Balance Reload feature is incredibly convenient for anyone who shops frequently on Amazon. It allows me to add funds directly to my Amazon account without having to go through the process of entering credit card information every time I make a purchase. This is perfect for those who prefer to manage their shopping budget by preloading funds to their account. It saves me time and makes the checkout process smoother.
Ease of Use
Reloading my gift card balance was quick and easy. All I had to do was go to my Amazon account, select the “Reload Your Balance” option, and choose the amount I wanted to add. The process is simple and user-friendly, with clear instructions. You can choose to set it as a one-time reload or opt for an automatic reload at regular intervals, which is an excellent feature for those who want to stay on top of their shopping without worrying about running out of funds.
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One of the best aspects of this feature is the flexibility in reloading amounts. I can reload any amount I choose, whether it’s a small amount to cover a single purchase or a larger amount for future shopping. There’s also the option to set up an auto-reload, so my balance is automatically replenished when it drops below a certain threshold. This is particularly helpful for frequent Amazon shoppers, as it ensures there’s always enough funds available without having to manually reload every time.
Processing Time
Reloading my balance was instantaneous, and I was able to make purchases immediately after adding funds. The money is applied to my Amazon account almost immediately, so there’s no waiting around for it to process. This makes it ideal for urgent purchases or spontaneous shopping.
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The Gift Card Balance Reload option is secure, and I have peace of mind knowing that my payment information is kept safe through Amazon’s encrypted systems. I don’t have to worry about storing credit card information for each transaction, and the reload process is secure and straightforward. I also appreciate that I get an email confirmation every time my balance is reloaded, providing an extra layer of security and transparency.
Value for Money
The Gift Card Balance Reload is a great option for managing your Amazon shopping budget. It doesn’t come with any additional fees or hidden costs, and I get exactly what I pay for. Plus, I can take advantage of any promotions or bonuses Amazon may offer for gift card reloads (e.g., occasional bonus offers or discounts). The ability to add funds as needed without being locked into a set amount of money is very useful and cost-effective.
Overall Impression
I’m highly satisfied with the Amazon.com Gift Card Balance Reload service. It’s a fast, easy, and secure way to manage your shopping budget and ensure that you never run out of funds on Amazon. The process is simple, and I love the flexibility in choosing how much to reload, as well as the option for automatic reloading. For anyone who frequently shops on Amazon or wants to manage their spending more effectively, this is a great tool to have.
I will definitely continue using this feature to make my Amazon purchases even more efficient and hassle-free.
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Reviewed in the United States on November 23, 2024