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Description
Mouse Kim1 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 conjugate are sequentially added to microwells pre-coated with Kidney Injury Molecule 1 (Kim1) 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 Kidney Injury Molecule 1 (Kim1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Kidney Injury Molecule 1 ELISA Kit；mouse Kim-1 elisa kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Kidney injury molecule 1 (Kim1), also known as hepatitis A virus cellular receptor 1 (HAVcr-1) or T cell immunoglobulin and mucin domain 1 (TIM-1), is a protein encoded by the HAVCR1 gene. It is a member of the TIM (T cell transmembrane, immunoglobulin, and mucin) gene family and plays a key role in regulating immune cell activity, particularly the host response to viral infection. It is also involved in allergic reactions, asthma, and transplant tolerance. The TIM gene is a type I cell surface glycoprotein composed of an N-terminal immunoglobulin (Ig)-like domain, a significant mucin domain, a transmembrane domain, and a short C-terminal cytoplasmic tail. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
Based on 2349 reviews
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Product Reviews
★★★★★ 5
Great book of fun facts about nation’s capital!
Format: Flexibound
I take my 3 boys (3, 5, 7) to Washington DC each year. This is a wonderful book full of fun facts for our nation’s capital. If you are looking for a kid version of a travel book that maps you through neighborhoods, etc., this is not it, but what kid would like that kind of book? That’s what grown-ups are for - mapping out the trip. Rather, this is a great supplement to read at bedtime to learn all sorts of facts about the city - from the historical pets of the White House to the error in the inscription on the Abraham Lincoln memorial. Really - these are great facts for adults also! Each page is a separate set of topics on its own, so it’s easy to read just a few pages at a time. Also there are great illustrations to hold the younger audience’s interest as well. This is a great buy and a must-have to get kids ready for their trip, or to read during it, or after (or all three!).
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Reviewed in the United States on May 28, 2018
★★★★★ 4
Happy Wanderer
Format: Flexibound
City Trails is not a guided walking tour (like the Freedom Trail here in Boston) of the Metro DC area. No addresses or street names are noted in the blurbs. To actually visit any of these places, you’ll have to consult a real map. For instance, the chapter “Statue City” highlights notable statuary around town. But the Capitol Building statues (in SE DC) are far from the Cathedral ones (in NW DC.) The themed groupings (G-G-G-Ghosts, Animals Around Town, Water World and more) are less maps to any place and more of an interesting overview of our Nation’s amazingly diverse and action-packed city. It’s best read as a primer on experiencing the flavor of the city (I lived and worked there.) It reads more along the lines of the “Weird But True” series made famous by National Geographic for Kids.
I don’t see this being of value to tourists in town for a limited time whose sightseeing is going to include major attractions like government buildings (White House, Capitol), museums (Smithsonian), some monuments (Jefferson, Lincoln, Washington) and other popular sites (Ford’s Theater.) This guide is actually best suited for the Metro-area (WDC, MD and VA) resident – child or adult - who wants a deeper dive into their hometown’s off-the-beaten-path sights and stories. A well designed and written book of historical trivia.
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Reviewed in the United States on August 29, 2018
★★★★★ 4
Nice way to learn about a trip to D.C.
Format: Flexibound
I got this for my kids to read before we went to Washington D.C. The pages are colorful, illustrated, and have short bursts of interesting details about the various attractions available to tourists who are visiting. My kids were eager to find the places on our itinerary and read about them ahead of time. They learned what to expect and were sure not to miss the important aspects of our tours. This book is recommended for 9 to 12 year olds and I think that is the perfect range. There is just enough information to peak their interest and not so much that they get bored by reading a bunch of text. The Table of Contents wasn't that informative in finding specific places, but the index was. My kids preferred to leaf through the whole book and find what was interesting to them.
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Reviewed in the United States on December 28, 2018
★★★★★ 5
Learning while having fun
Format: Flexibound
Great book for the grandchildren - and the parents enjoyed it with them
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Reviewed in the United States on January 20, 2018
★★★★★ 3
Not good for travel, picture book only
Format: Flexibound
This is a beautiful book with fantastic illustrations and an engaging color palette. It includes a good variety of historical background information and sightseeing locations. However, it is a better picture book for browsing only. It is not a good travel book for kids to plan their own adventures. Who has ever heard of a travel book without maps?! No maps, no directions, no coordinating subway/bus maps. The printing is exceptionally small, almost too small to read. The book should have been made larger to accommodate the text. The text is excellent, but it printed as if the publisher never expected the kid/parent to read it. On one hand, our family really enjoyed how engaging the book appeared, but we were disappointed in its quality as a travel book. It shouldn’t be marketed as a travel book, but a geography book series.
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Reviewed in the United States on February 1, 2020