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Description
Human MMP-12 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 enzyme conjugate are sequentially added to microwells pre-coated with a matrix metalloproteinase 12 (MMP-12) 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 matrix metalloproteinase 12 (MMP-12) 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 12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Matrix metalloproteinase-12 (MMP-12), also known as macrophage metalloelastase (MME) or macrophage elastase (ME), is an enzyme encoded by the MMP12 gene in humans. This enzyme degrades both soluble and insoluble elastin. This gene is part of the MMP gene cluster located on chromosome 11q22.3. Proteins in 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. Most MMPs are secreted as inactive proproteins. When the enzyme is activated, the prodomain is cleaved by extracellular proteases. The active enzyme consists of two domains: a catalytic domain responsible for its enzymatic activity and a hemoglobin-like domain, which, in some MMPs, plays a role in substrate recognition and contributes to catalytic efficiency. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 4
Looks great!
My son loves these shelves. He wants more!
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Reviewed in the United States on October 9, 2025
★★★★★ 5
Husband loved it!!
Got this for my husband on Christmas and he absolutely loves this. The instructions were straightforward on how to build and it took less than an hour to assemble. The lights are pretty cool. They change different colors and there's different settings with the remote. I will say he did have to nail these to the studs in the wall so it can hold his hefty drinks LOL
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Reviewed in the United States on January 29, 2025
★★★★★ 3
A royal pain to install
The width of this item is such that it CANNOT be attached to the wall studs. That is unsafe for something that is intended to hold heavy items like liquor bottles. THEN the anchors that come with it are flimsy so we used a heavier different style anchor (threaded). That said, I like the way it looks, and the light effects.
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Reviewed in the United States on January 4, 2025
★★★★★ 5
New Bourbon Shelf
I just got done putting it together and hanging it on my wall. It took approximately 35 minutes to put together and another 15 minutes to hang up. I had to use my own larger wall anchors and screws. I tried the ones that came with it but they didn’t do the job. I am happy with it other than that. Good value and quality materials. As you can see from the picture it holds several bottles and room for more if I want. I think the LED lights look great as well.
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Reviewed in the United States on April 18, 2025
★★★★★ 5
Mocks dispensational viewpoints on key Thessalonian chapters.
Format: Hardcover
Very good read. You really walk away feeling like an expert in the Thessalonian Epistles after reading this book because Weima always is searching for theological answers to the questions that arise within the Epistles themselves. You really see the deep unity of Paul's purpose in writing these epistles after reading this commentary. The only problems for me where I did not agree with Weima's conclusions was on dispensational issues in 1 Thess. 4 and 2 Thess. 2. Weima rejected the idea of the rapture without dealing with it at all and then proposed the idea that its talking about a "Hellenistic reception" of a king coming into a city. I don't get it, Weima thinks we do go to the sky meet the Lord in the air and then lead him to the earth, it's so preposterous. It seems like God is in control of this event not man leading Christ to the earth. If he wants to criticize the rapture 'which yes I do believe.' Make your argument. But instead Weima just says things like "so called rapture" without refuting it with logic at all. And then in another eschatological text in 2 Thess. 2 Weima rejects any kind of a literal interpretation and says "We can't actually believe that the Anti-Christ actually goes into a rebuilt Jewish temple as some dispensationalists believe." I'm paraphrasing Weima's words. Weima flat out rejects any dispensational viewpoints on these important verses and doesn't deal with them at all like he does other viewpoints. Other than that I think this was a great read. Highly recommend the book.
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Reviewed in the United States on February 28, 2016