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Description
Mouse MMP12 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50ng/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 12 (MMP12) 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 12 (MMP12) 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 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. 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 may contribute 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 374 reviews
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Product Reviews
★★★★★ 5
What's to say, a Saint and a Doctor of the church.
Format: Paperback
Content awesome as one would expect from a doctor of the church. Problem, font size of the printed is to small for comfortable reading.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 17, 2025
★★★★★ 5
A guide for prayer relevant today
Format: Paperback
Though written in the 16th century, this book is so direct, speaking of the issues of parenting, prayer, and our relationship with God, as if the author were alive today. The examples she gives are so human, and so direct. Do you pray in church, or do you fall into the temptation of looking who is there around you? What other temptations do you give into that take you AWAY from prayer? Do you know how easy it is to falter in prayer? St. Teresa will help you not feel like a failure. There are times you should simply not pray ("no water in the well"), and in those cases, St. Teresa suggests to read a good book. Lots of good books - and hers is a perfect way to start. She is helping me pray, but when I am having a dry spell, she is helping me not fall away from God.
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Reviewed in the United States on April 22, 2014
★★★★★ 5
......................COINCIDENCE?…I was on page 130 when my husband
of 43 yrs kissed me good by, walked out the door, fell down -& never got up. His last words to me when I tried to help him up were " I'm ok".
The paramedics worked on him for over an hour but never got a pulse. He died less than 5' from me -but, I couldn't even hold his hand.
He'd converted to Catholism but, became a Catholic without a church; like me!
Why I chose this book is still beyond me......especially of this Saint; we'd been to her shrine, in Spain, yrs ago. I'd found it
'too much'… the relics (a finger) -as I remember it struck me as wrong.
On that same trip, we went to Fatima; my husband told me he just knew something special had happened here when we visited the shrine & the small statue of Our Lady of Fatima. I,
(the Catholic, raised by devote parents & nuns in school) who really wanted to feel the same -didn't.
It is 3wks. since his death.
Maybe, I'm just still numb.
We were 24/7 for over 43 yrs. (we had a business- then retired & traveled - we had a wonderful life) BUT…
How could I live now...without him? And Why?
I waited for a 'breakdown'… for his loss to hit me. I felt waves of grief, that were physical -go through me for a few days; but, no great sobbing of tears.
The only explanation I have (& I thank God for it) is that I do feel him with me still, in my heart/soul. My daily prayer is that I never feel That loss. (being the cynic that I am, I can not begin to express what this means to me) It is so real a presence & comfort!
And, 'for the record', the very first call I made was to the church we weren't going to. (In 'my' day, if you weren't in 'good standing' you didn't get a funeral Mass.) We got a it and were welcomed!
My Faith, is still pathetically weak. Father told me to offer my grief as sacrifice. I'm trying.
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Reviewed in the United States on November 25, 2018
★★★★★ 5
Very thoughtful, encouraging, and personal reflection of her life
St. Teresa’s personal reflection of her life brought me a clearer light and understanding of her other more theological writings. What impressed me most was how she revealed her own many short comings and failures despite the great graces she received. This was a real encouragement to me in my own journey toward becoming a saint.
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Reviewed in the United States on January 28, 2022
★★★★★ 4
Illustrated ?
The content of the book is well worth reading. The claim that it is "illustrated" borders on dishonesty. There are 14 thumbnail photos, none of which have anything to do with the life of Teresa of Avila. However, it was worth $0.99 (for the Kindle version).
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Reviewed in the United States on April 23, 2019