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Description
Human TIMP-2 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 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 Tissue Inhibitors of Metalloproteinase 2 (TIMP-2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Tissue Inhibitors of Metalloproteinase 2 (TIMP-2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tissue Inhibitors Of Metalloproteinase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Matrix metalloproteinase inhibitor 2 (TIMP-2) is a gene and a corresponding protein. This gene is a member of the TIMP gene family. The proteins encoded by this gene family are natural inhibitors of matrix metalloproteinases (MMPs), a group of peptidases involved in extracellular matrix degradation. This encoded protein is crucial for maintaining tissue homeostasis by inhibiting the proliferation of angiogenic factors in quiescent tissues and suppressing protease activity in tissues undergoing extracellular matrix remodeling. TIMPs inhibit active MMPs, but different TIMPs have different inhibitory effects on different MMPs. For example, TIMP-1 inhibits MMP-7, MMP-9, MMP-1, and MMP-3 more effectively than TIMP-2, while TIMP-2 inhibits MMP-2 more effectively than other TIMPs. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenates and other biological fluids |
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4.2 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 5
beautiful story
Format: Kindle
As always a treat for your imagination. Beautiful people with even more beautiful souls. This was a great short story and a lovely quick read with a happy ever after. Thank you for writing such a delightful story. If it was a 1000 pages I would have loved it just as much. Great job…
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Reviewed in the United States on March 23, 2025
★★★★★ 5
So much feelings!
Format: Kindle
Collette was not voiceless in the least and to say she is by far my fave FMC of all time right now it's no understatement. Best 1st read of the year ever!! I love how possessive each of her men were and OMG Lena is the best BFF and girl could ask for! I'm pretty sure I need this gorgeous book on my shelf pronto!
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Reviewed in the United States on February 1, 2025
★★★★★ 5
A Fox Amongst the Wolves
Format: Kindle
Collette has been forgotten about her whole life. Her parents don't bother with her, others don't try either once they realize she uses sign language to speak, and she's a shifter surrounded by humans that hate her kind. On her 18th birthday, she runs away. She takes the train and then bus wherever it can get her as far from home as possible. A missed bus brings her and a mysterious but kind man to Willowdale. What happens next is a story of fated mates, shifters, magic, new best friends and found family, and a stolen child. I'm sad this was a shorter read but only because of how much I loved Collette, Dylan, Hunter, Luca, Lena, and the others in their community. I love how detailed the characters are even with the shorter story. You can tell their characteristics easily, and the lead in to book two is done so well. I loved this insta love book with just the right amount of drama.
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Reviewed in the United States on July 10, 2025
★★★★★ 4
An easy PNR book to cure book hangovers.....
Format: Kindle
I enjoyed this as a short story; but nothing truly pulled me into this world to want to keep reading each story. I can sense the overarching villain arc w/ the HAF; yet it still wasn't enough to capture me long-term. It felt like too much focus was placed on progressing the romance part that everything else took a non-existent back seat. This did work well for my intentions of getting over a book-hangover slump. So there's that lol. A Tip to overcome a PNR book hangover, from a wonderful series or novel......read a trope-y, lighthearted, PNR that is easily predictable but adorable. Like this book.
Honestly, I'm rarely into short stories that are turned into a ridiculous amount of books on different PNR characters. So I'm not the best at judging the merits of these stories. I'm a long-haul type of reader that lives for a series surrounding 1 lead female character & her harem (or singular partner). Occassionally, I'll find a series of short stories that are interesting enough to finish or pull me in completely. The rarity of that is countable on 1.5 hands, compared to the literal hundreds of fiction books I've read lol.
With all of that, I will say this was fun because of the FMC's difficulties growing up & how that made her strong, the end reveals of her life, & the cute Epilogues. I never felt truly connected to any of the characters; but I did feel slightly compelled to just finish it to see how plot points would end.
Unfortunately, due to a lack of character backstory explanations, I still don't understand Collete's "magic". Very little time was given to flesh out the backstory of the world this occurs in. I couldn't even guess what type of magic occurs in this world, aside from what little was revealed. A random scene sticks out to me...... we're introduced to a new magic type, in passing. No dialogue to expand. No hint at anything really. Then the scene moves on. That moment felt pointless to even mention, afterwards.
It felt like the author was more concerned w/ romance than storyline. This is a common issue I have w/ short stories....authors write them for whatever reason; but too often it's an "either or" theme.....EITHER the author is writing ONLY for romance w/ storyline taking a minor role OR writing ONLY for storyline w/ some romance that ends up feeling forced. As I mentioned earlier, only a handful of short stories series have ever had a balance of both sides, for me, that I wanted to read all of them.
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Reviewed in the United States on May 16, 2021
★★★★★ 5
Great RH shapeshifting romance!
Format: Kindle
Oh my gosh, I adored this book! Voiceless is set in a blended reality with humans, witches, and shapeshifters of all kinds, but with a significant lack of acceptance from the humans that results in violence. And I am sucker for fated-mates/insta-connection and you get that in heaps and bounds in Voiceless! Colette’s introduction to the supernatural world added some challenges on top of not being able to effectively communicate with those around her. I enjoyed the overall plot; that there was danger and confrontation and resolution, albeit not nearly as much as I wanted. Voiceless is a short read and things escalate at a fast pace and a resolution is reached just as quickly. As a result, I think we miss out a bit on relationship and character development that we could’ve had if the book was little longer. While this is a RH/why-choose, most, if not all of the spice, is exclusively MF.
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Reviewed in the United States on March 17, 2025
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