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Description
Mouse VTN 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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 400 ng/mL). Then dilute to the following concentrations: 400 ng/mL, 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 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 400 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 200 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 vitronectin (VTN) 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 vitronectin (VTN) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Vitronectin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| 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 | 6.25-400 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.3 ★★★★★
Based on 614 reviews
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Product Reviews
★★★★★ 1
A Catastrophic Failure in Diaper Form
Size: Size 2 (184 Count)
I’m not sure if I just got a bad batch or if these diapers are always this horrendous, but I will not spend another dime to find out—and I strongly urge you to do the same. I loved this brand’s Gentle version. They were everything you want in a diaper: super absorbent, well-fitted, no blowouts. I was their biggest fan. So, when the Gentles weren’t available, I thought, Surely the Plush will be just as good, maybe even better!
I have never been more wrong.
Let’s start with the tabs. These things are folded up like some kind of sick joke, requiring you to unfold them in three separate, precise movements. Miss a step? The tab rips off in your hand, leaving you holding a now-worthless scrap of disappointment. This happened to me. It happened to my mother. It will happen to you.
But wait, there’s more! The side wings are a disaster. The right wing is disproportionately longer than the left, which means everything in this diaper is permanently listing to the right like a sinking ship. The left side? Practically non-existent. As a result, I have experienced more blowouts with these diapers than in my daughter’s entire life combined. I don’t know what sorcery was used to engineer a diaper that actively funnels poop toward the worst possible escape route, but here we are.
And because I was foolishly optimistic, I bought a month’s supply. So now I'm locked in a psychological battle with my own stubbornness. I could throw them out, but that would be admitting defeat. So instead, I soldier on, knowing each diaper change is another step into madness.
Please, learn from my mistake. You don’t have to live like this. Let me be your tribute, the sacrificial parent who suffered so you don’t have to. We’re already running on fumes as newborn parents—don’t make it worse by willingly subjecting yourself to these truly useless diapers that will rob you of your last remaining shreds of sanity.
Save yourself. Run.
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Reviewed in the United States on February 26, 2025
★★★★★ 5
DON'T SLEEP ON THESE DIAPERS! (but your baby should)
Size: Size 3 (40 Count)
I was skeptical at first trying out the plush version of this diaper because I thought the mama bear gentle touch was just OK and you can find a better diaper for the same price point. I saw these on prime day for the same price as the gentle touch and said why not. I was very, very impressed. They are soft and flexible with a satin-like exterior; similar to the feel of a hello bello diaper. The plush version definitely feels more quality than the papery gentle touch ones. I first tested the absorbency using 12 oz of water, waited ten minutes and dabbed dabbed dabbed. Seriously NO moisture! In comparison to the gentle touch diaper that had obvious moisture after the same test. I put them to use on my baby, who just for reference is 19lbs, for an overnight and the diaper did not leak and was only damp in the morning, not wet! Amazing. She is a moderate wetter but I still think that performance is great because in comparison to gentle touch, the diaper was wet on the inside after only three hours and I had to consistently use diaper cream so rash wouldn't develop; I would have never used the gentle touch as an overnight diaper. Seriously, for .21 a diaper for size three these diapers are a steal. DONT SLEEP ON THIS DEAL, PICK UP SOME MAMA BEAR PLUSH DIAPERS!
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Reviewed in the United States on July 23, 2024
★★★★★ 3
Not the Same 🤍
Size: Size 2 (184 Count)
I loved the Mama Bear size 1 diapers, so I expected the “plush” size 2s to be the same. Unfortunately, they feel completely different. These seem thinner and not quite as absorbent as the size 1 regular pack.
The design is also really busy, which makes the wetness indicator harder to see at a quick glance. With babies, that little line matters more than you’d think.
I’ll still finish the pack, but I’m not sure I’d repurchase this “plush” version. If you’re choosing between them, I’d recommend sticking with the regular pack instead. In comparison, Huggies still perform better than these for us. 🤍✨
They don’t hold blowouts at all but they do fit well. They’re also not stretchy. I noticed the print on my baby’s legs.
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Reviewed in the United States on March 14, 2026
★★★★★ 5
Great cost efficient option
Size: Size 4 (36 Count)
I switched over from Millie Moon and really like these. They’re affordable, don’t have harmful additives, and are soft and gentle on my babies. Holds a significant amount and doesn’t leak even on overnight wears. I like the simple non character design as well.
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Reviewed in the United States on February 2, 2026
★★★★★ 2
Every other diaper leaked
Size: Size 7 (92 Count)
I bought these diapers because they were the cheapest available to buy with a gift card my family was given. Normally I use the bulk store brands. The diapers were very hit and miss. It varies from diaper to diaper even in a pack. My son does urinate heavily but even before he would get anywhere near a “full” diaper it would leak! Leading to wet pants and messes. We’ve never had this problem with other diaper brands. Even our cloth diapers hold better than half of these did. For note he falls below the max size for these diapers by at least 5 lbs. I don’t plan on repurchasing these because of the leaking issue. Beyond that they were easy to use like any other diaper, didn’t cause diaper rash, held together, but was subpar in absorption.
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Reviewed in the United States on November 19, 2025