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Description
Rat 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. 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. 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. 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 | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Tissue Inhibitors Of Metalloproteinase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tissue inhibitor of metalloproteinases 2 (TIMP2) is a gene and corresponding protein. This gene is a member of the TIMP gene family. This protein is considered a metastasis suppressor. It is a specific inhibitor of MMP-2, binding to activated or inactive MMP-2, terminating MMP-2 activity and effectively inhibiting MMP-2's collagen and gelatin degradation activities, thereby suppressing cancer cell metastasis and invasion. Numerous studies have shown that TIMP2 and TIMP-2 are two indicators of tumor invasiveness, playing a crucial role in the metabolic balance between extracellular matrix synthesis and degradation. Disruption of the balance between MMP-2 and TIMP-2 in tumors is a key factor in tumor progression. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 3
Not Durable…Splintered….
Had to chuck these. I loved them at first but they are not durable. Scratched themselves up right away and then the mid sized one started to splinter. I didn’t even have it for very long before I had to get rid of them. Long enough to not be able to return. I prefer the bamboo as far as wood goes, but switched to an antibacterial. It’s crazy because I had the same cutting boards for years and in the past month I’ve had to replace them 3 times now. I was so disappointed because there were such amazing reviews. Sadly that was not the case for me. I give it a 3 because they are absolutely beautiful…..you get points for being beautiful I suppose in this world. But not durable. At least not for me.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 22, 2026
★★★★★ 5
Perfect cutting board set of 3
Very nice cutting boards, and I especially love the peace of mind knowing there is not harsh chemicals used to make these. Thank you!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 27, 2026
★★★★★ 5
Fascinating exploration of God's Word!
Urban Legends of the New Testament: 40 Common Misperceptions is an impressive work by David A. Croteau that discusses 40 "commonly circulated myths, repeated throughout the culture as common knowledge, but which isn't true."
But we're talking about the infallible Word of God, right? That is correct. But according to the author, certain passages of the New Testament have become victims of the notorious Urban Legend phenomena.
The titles of each chapter are the actual legends and are presented as if the author really believes what he is writing to be true. The reader will soon find out that is not the case. Croteau goes on to discuss the incorrect interpretation versus an accurate interpretation and then ends the chapter with his opinion of what the text means, followed by a practical application section.
The presentation and discussion of each myth is clear, although some are harder to piece through than others. The length of the chapters vary according to the history of the myth and depth of the discussion. Topics vary from the birth, life, and crucifixion of Jesus to Paul's profession, the plan of salvation, hell, divorce, money, pastors and their families, and cults. You can pick up the book and read only the chapters that interest you, although I do encourage you to read the introduction and epilogue.
The prologue discusses two types of legends: a mistaken-contains wrong information; misleading-those that need clarification. The cause and solution for urban legends of the New Testament come down to three components: Context, Greek, and Backgrounds. Croteau clearly discusses this at the end of the book and my only criticism is that I would have liked that information in the introduction. I believe it would have aided in my overall structure for comprehension.
Overall, the research was fascinating and I learned—a lot.
This quote from the final conclusion summarizes the purpose of this book:
"In the end our goal should be to interpret Scripture as accurately as possible, to apply that interpretation to our own lives, and to be transformed by God's Word so that our lives may bring glory to the One who rescued us from an eternal destiny in hell."
I highly recommend this book to all readers. Don't plan to read it in one sitting. Use it as a study guide and tool to inspire you to dig deeper into God's Word.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 12, 2016
★★★★★ 5
Only Orthodoxy Will Lead to Orthopraxy and Orthopathy
Format: Kindle
This book authored by David Croteau is truly an enjoyable yet at the same time a convicting read. Even though it was written about 10 years ago, the principles of interpretation advocated by him to deal with the "mistaken and misleading legends" are timeless. Out of the 40 legends, I have encountered at least half of them in the local church, writings and even seminary especially issues pertaining to salvation, Christian living and women's adornment.
The main takeaway is this: Just as there are three things that matter in property: "Location, Location, Location", similarly the three main things that matter in interpretation of the Word is: "Context, Greek, and Background Information", or even simpler: "Context, Context, Context". David Croteau has demonstrated in his 40 legends the utmost importance of context in correctly interpreting a particular passage which was unfortunately taken out of context. The interpretation can further be handled even more accurately by taking into account the Greek language and the socio-historical background behind the text. In fact, I have been focusing on these three elements in my hermeneutical approach whenever I come across passages which I need more understanding. I don't know Greek; but with the many useful online Greek tools nowadays, I can always do a word study which more or less balances my lacking of Greek knowledge. Also, the study of socio-historical background is extremely illuminating if one have access to these resources. Personally, I discover that a proper understanding of background information can eliminate most of the mistaken or misleading interpretation of a text.
Lastly, I totally agree with David Croteau's conclusion: "In the end our goal should be to interpret Scripture as accurately as possible, to apply that interpretation to our own lives, and to be transformed by God’s Word so that our lives may bring glory to the One who rescued us from an eternal destiny in hell." His conclusion reminds me of a phrase which I encountered years ago: "Orthodoxy Will Lead to Orthopraxy and Orthopathy." Indeed, only by "accurately handling the word of truth" (2 Tim 2:15) can we realise the necessity of holy living and the proper response that we should have towards our living God.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 6, 2025
★★★★★ 5
The book is also great in pointing to additional resources for further study
Format: Paperback
Other than the Bible, I hesitate to use the term "must read" for a book. This one is definitely a "should read" for anyone who teaches, preaches, or wants to minister more faithfully. The book is engaging, and offers both a negative (doesn't mean this) and positive (means this) response to common urban legends of the NT. You'll be surprised by some beliefs you had that you didn't even realize were not correct, or should be adjusted. The book is also great in pointing to additional resources for further study. Pastors, seminarians, and lay people will all benefit from this book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 17, 2015
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