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Description
Rat ZNT1 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 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 a capture antibody against zinc transporter 1 (ZNT1). 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 zinc transporter 1 (ZNT1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Zinc transporter 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Solute carrier family 30, member 1 (SLC30A1), also known as zinc transporter 1 (ZNT1), is a protein encoded by the SLC30A1 gene. It regulates zinc permeation through L-type calcium channels. It downregulates not only Zn++ influx but also Ca++ influx, thereby protecting cells from excessive cation permeation. It may be involved in zinc transport out of cells. Diseases associated with it include zinc-deficiency acrodermatitis enteropathica and epidermodysplasia verruciformis type 1. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
A great toy for any dog!
Style: Small Ball, Size: 1 Count, Style: Small Ball, Size: 1 Count
The Hartz Duraplay ball is great! My 4 1/2 yr. old Pug has never liked playing with them before. He has gone crazy for this ball. He has no trouble grabbing it with is mouth. He ever gets it to squeak with his feet. I think it’s a great dog toy. Excellent!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
★★★★★ 5
Great Value and very durable
Style: Small Ball, Size: 1 Count
Very durable! We have purchased this for our new lab puppy. Highly recommend them. We have bought them before for our retrievers and it has been one of the longest lasting toys they own. Which says a lot because toys get torn up fast in our house. Definitely a good value and the material is easy on their teeth. Also it keeps ours busy for a while.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2026
★★★★★ 5
Great product! Durable, chewable, awesomeness!
Color: Grilled color, Color: Grilled color
Very well made and durable. My dog hasn't left them alone since i opened them. Hes been at it for about 45 minutes straight now! Lol. He's an avid chewer. He will chew thru and fabric or cloth toys in less then a day. But these are what he really likes because they take time to chew apart. The shapes, are great, because one he props up with his paws and chews and it seems like it's easy to grip for him. Thru are hard so if you have hard floors it can get a bit noisy when he drops them. Over all this is a great product. Safe, chewable, long lasting, fun shape looks real to him (i think lol), Me and my dog Lucky are both happy with his new toys!
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Reviewed in the United States on March 27, 2026
★★★★★ 5
Recommend for very heavy chewers
Color: Grilled color
My aggressive chewers love these! It’s been a few weeks and still in one piece and man they chew them for hours on end.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 26, 2026
★★★★★ 5
Amazingly great product!
Color: Grilled color
I’m not just saying this, ok, this is not hyperbole: this is THE best nylon bone for chewing that I have ever purchased. And you get TWO of them for relatively less expensive than most chew toys give you one! I have 2 golden retrievers, and if you know goldens, you know that they are some of the most relentless and powerful chewers around. So I can’t give them almost any toy other than rubber stuff or it will be destroyed in a period of time that doesn’t make it worth it. One of them even figured out how to destroy a supposedly indestructible toy made out of kevlar! She was able to get into it through the seam, which is not kevlar ha ha. Anyway, I digress; I never even considered nylon chews because they suck! Usually. Usually my dogs in the past have lost interest with them in less than a day. I took a chance on these mainly because of the novel shapes, the fact that they give you two, and the low price. Both my goldens LOVE these! Not just for one day, but going on weeks now! I don’t know if it’s the flavor or what, but I will see them gnawing on one of them at some point every day. And they CAN’T destroy them (I knew nylon was good for that, but it was the “who cares” attitude of my previous dogs toward nylon that kept me away). Both the “ribs” and “steak” (quite clever) are a bit worn on the edges now, but they look like they will be around for years. You can’t go wrong with this product (unless for some reason your dog does not like the flavor, but the flavor seems to be what draws my dogs to them). I highly recommend this (these) as the most cost-effective chew toy I’ve ever purchased.
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Reviewed in the United States on November 1, 2025
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