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Description
Mouse MMP13 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 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. 3. 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 13 (MMP13) 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 13 (MMP13) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Matrix Metalloproteinase 13 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Matrix metalloproteinase 13 (MMP-13), also known as collagenase 3 (GLG3), is an enzyme encoded by the MMP13 gene. It is a member of the matrix metalloproteinase family. Like most MMPs, it is secreted as an inactive proform. Once the prodomain is cleaved, it becomes activated, leaving the active enzyme consisting of a catalytic domain and a hemopexin-like domain (PDB): 1PEX. Although the exact mechanism remains undetermined, the hemopexin domain is involved in collagen degradation, while the catalytic domain itself is particularly inefficient at collagen degradation. During embryonic development, it is expressed in bone, where it is required for the reorganization of the collagen matrix for bone mineralization. In pathological conditions, it is highly overexpressed. Its transcriptional regulation is tightly controlled due to its efficient proteolytic capacity. Several cytokines and growth factors have been shown to influence MMP13 gene expression, including parathyroid hormone, IGF-1, TGF-β, and hepatocyte growth factor. | |||||||||||||||||||||||||||||||||
| 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 | 31.2-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.9 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 4
Decent fit, good quality, great value
I'm 6' 4" tall (1.93m), but very thin, so I bought a Small, which is the size I wear for almost all men's short-sleeve shirts (tees, polos, and button-downs). The fit of this shirt is surprisingly roomy, although I'm not complaining given the price point. The shoulders and body could be a just tiny bit more tailored, especially given the easy drape and movement of this pique knit polyester fabric, which hangs where it wants to and glides effortlessly against the skin. I could never, ever feel constrained or constricted in this shirt. I almost wish the fabric were a tiny bit stiffer. The length is slightly generous, but not excessive. The collar lies down properly (no curling at the corners, no unwanted creases), although this is usually the case with synthetic fabric polos. Despite the minor flaws with this shirt, I like the overall appearance, value, and quality, and I plan to buy several more in different colors.
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Reviewed in the United States on August 13, 2018
★★★★★ 5
A shockingly good polo shirt!
OK. Amazon Brand. How good can it be? Well, shame on me. I bought one as "cheap" knock around polo shirts for the beach However, what I received was actually a very nice quality polo shirt with a great fit. (Large, "Trim fit" fits me well....average build, 5'9", 155 lbs, 32 waist). While I can appreciate nice clothes, I am only as fashion conscious as the average 58 year old male. With that caveat, I would unquestionably wear this golfing, at summer party or at a causal restaurant . The material has a nice feel, dries quickly and looks like it will resist snagging, though that it a risk with any polyester shirt. I cannot yet speak about long term durability, but I will no doubt recover my $15 investment. I have since ordered 5 shirts more because I lost weight and needed new shirts. Is this shirt as nice at the $90 ones my wife buys me at a "high end", unnamed "preppy" shop? No, not quite. But awfully close!
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Reviewed in the United States on June 23, 2021
★★★★★ 5
Buy this shirt!
You need to buy this shirt! I bought one for a golf outing and like it so much I went back and bought all the colors that were available. At $15, these are a superb value and look great.
Fit: this is an athletic / slim fit shirt. I am 5’9” and 160 lbs and a Medium fits me perfectly. Fits much better than Banana Republic polos. The collar sits nicely, the button placement is correct (not too high not too low). The cuffs do not have a band so they don’t ride up.
Fabric: 100% polyester so you can wash it and stick it right in the dryer and it comes out with no wrinkles. NOTE: I had trouble ironing this shirt. Best bet is toss it in the dryer for a few minutes to get wrinkles out. The colors I have are vibrant and there is a slight texture but not as much as a pique.
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Reviewed in the United States on September 6, 2019
★★★★★ 3
Nice shirt but too short
I'm 6', 175 with a bit of a longer torso. That makes me a slim build bordering on tall/slim. The material seems on par with similar offerings from Nike and the shirt looks and fits quite well (I like the slim tailor) but it was not long enough to tuck in as a medium (right at my belt line). I tried a large which is getting a little big around and it was still too short. XL was ridiculously wide on me and still not long enough.
Bottom line is that Amazon needs to offer this shirt in a slim/TALL with about 3" of additional length. There would be a whole lot of buyers. I'll have to continue to go to the specialty shops in search of MT and LT sizes.
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Reviewed in the United States on July 31, 2021
★★★★★ 5
Nice for the price
I am 6’4” and 195 lbs so I need a large size for most shirts. I need it for the length, but not the width, so shirts this size tend to bow out at the back and make me feel a bit like I am wearing a tent. I prefer polos made of a very thin, breathable and moisture wicking fabric. A decent looking shirt you can wear on hot and humid days without any undershirt and stay comfortable. Old Navy makes a fantastic version, very similar to the ridiculously priced Under Armor version, and I was hopeful that this Amazon version would compare. It does, but has a few drawbacks. If this sounds like you, keep reading. If not, thanks for getting this far.
I’ll keep this brief with a list of pros and cons below. Suffice it to say, I like the polo enough not to return it, but it is still a step below some of the shirts produced by other outlets.
Cons:
1. It is far from wrinkle free, though I’m not sure it is even touted to be. It does tell me, though, that this fabric won’t be as soft as I prefer.
2. The color is a little darker (I got the coral) than it seemed in the picture, but not terribly so.
3. The “slim fit” is what drew me in and while it better than most, the back still bows out a bit
4. The fabric is clearly moisture wicking, but not the light fluffy kind. I know I’m starting to sound like a snob, but if you know what I mean, you know what I mean. Sometimes a shirt feels heavy when you sweat in it. The idea of the moisture wicking fabric is to pull that sweat away as fast as you produce it to aid in the evaporative cooling taking place on your body. This shirt doesn’t feel heavy, but I’m not sure how long it will stay that way on a really hot and humid day.
Pros:
1. Price. For $15, I can become blissfully ignorant to the cons listed above.
2. Fit. It isn’t perfect (Yes, I know this is listed as a con too), but if you have a tall, lanky body type like me, a large will fit you better than your average polo. After a trip or two through the dryer, probably even more so.
3. Convenience. Now that I know this is a good fit for me, I can try all the different colors I want from the comfort of wherever I choose to be with my phone.
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Reviewed in the United States on May 30, 2020