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Description
Human IFIT2 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 conjugate are sequentially added to microwells pre-coated with an interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) 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 interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Interferon-induced protein with tetratricopeptide repeats 2 | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interferon-induced protein 2 (IFIT2) is a protein encoded by the IFIT2 gene. It may play a role in preventing tumor progression. Indeed, IFIT2 expression has been detected in human papillomavirus-positive keratinocytes from cervical precancerous lesions, where it is gradually downregulated at different malignant stages. Therefore, this gene may be involved in tumorigenesis and may serve as a potential prognostic marker for the progression of cervical precancerous lesions. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Super tough, great value, addictive toy for my dog.
Flavor Name: Bacon, Size: X-Large
My dog pit/Shepard mix, loves this bone, and I love it because she can't kill this one like all those other Kong toys. She had a smaller nylabone, she killed it but took about a year, this larger one, she won't be able too that soon. Hard expensive Kong things, wouldn't last a couple months with her. So now, 3/17/2026 its still going. One end is gone, but the green part and other bone end she's working on.
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Reviewed in the United States on May 2, 2025
★★★★★ 5
Great for Mega Chewer
Flavor Name: Bacon, Size: X-Large
Got one of these for our Rodesien Ridgeback when he was a puppy, and it worked well. So, when my daughter rescued two medium-sized dogs (one is an Australian Cow dog mix, and the other a Plott Hound mix) it was a no-brainer. The Plott Hound mix has jaws of steel and destroys anything and everything he can get his mouth around, except items from this maker. This is the only toy brand I've found that he doesn't demolish in under 5 minutes. Oh, and I'm stunned he can put bite marks in it. The Rodesien Ridgeback didn't even do that when he was a puppy, so I was surprised. However, that being said, the toy is holding up to hours of abuse. I had to buy additional toys because they don't like to share.
Update on 01/09/20: these are still holding up to repeated abuse.
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Reviewed in the United States on September 21, 2019
★★★★★ 4
Sensational Toy By Nylabone Keeps Your Best Freinds Teeth Snow White!
Flavor Name: Bacon, Size: X-Large
Our Bull dogs LOVE these bones. Long time Nylabone fans they've been making quality dog toys here in the United States for over fifty years.
The nubs on the end of the bone age great for cleaning teeth and massaging bones. The center is softer but still holds up to even the toughest aggressive chewers. For whatever reason this isn't a "favorite" toy in the sense that they drag it around a few hours a day but they do play with it and we replace them when the ends start to shred - which should be done with all nylabone chew toys.
The price on Amazon is virtually half the cost of the big box pet stores so it's affordable - not to mention fun to replace their toys and try new ones often. Plus with a Prime membership almost every item we buy ships with Free two day shipping - making the deals even that much sweeter.
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Reviewed in the United States on December 16, 2012
★★★★★ 5
You've got to be kidding. Really !!!
Flavor Name: Bacon, Size: X-Large
I've used these for over fifteen years They are great. But, mine only lasted two days this time. Lucy the Labrador, left it in the driveway one afternoon. The next morning a huge garbage truck ran over it, making dust out of it. Normally, Lucy cant go any ware without it, so leaving it outside was her fault. And mine, for not checking. Yes, I ordered another one right away. Otherwise, she would have drove me crazy. These are great, and they last a long time, even with the most aggressive chewers. Lucy, passed away on June 16, at age 13, but you can bet I still have her favorite bone. The more a dog chews on these, the better they brush the dogs teeth. A+ product, just like their other items.
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Reviewed in the United States on June 30, 2015
★★★★★ 5
Large, Heavy, DURABLE!
Flavor Name: Bacon, Size: Medium
This bone is probably too big and heavy for my medium size cocker spaniel. But I bought it anyway, because he was able to demolish the Nylabones that were labeled "for medium dogs".
He can still pick this one up and carry it around, but he cannot fit an entire end into his mouth. Still, he loves it--he's greedy and relishes the fact that he has such a big bone all for himself. He even sleeps with it!
I like it because of the fact that it doesn't completely fit into his mouth. Due to this, the bone has lasted much longer than all the other Nylabones I bought him in the past. And because he can't shred this one, I don't find sharp bit of Nylabone fragments everywhere and it doesn't make his gums bleed either.
I happy I found this and will continue buying oversized Nylabone toys for my dog!
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Reviewed in the United States on March 21, 2012