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Description
Mouse SLC30A4 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 utilizes 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 4 (SLC30A4). 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 4 (SLC30A4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Zinc transporter 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Solute carrier family 30, member 4 (SLC30A4), also known as zinc transporter 4 (ZNT4), is a protein encoded by the SLC30A4 gene. It belongs to the ZNT family of zinc transporters. ZNTs are involved in transporting zinc out of the cytoplasm and share a similar structure, consisting of six transmembrane domains and a histidine-rich cytoplasmic loop. ZnT4 supplies zinc to zinc-dependent proteins in the trans-Golgi network, which is crucial for cellular function and zinc export in mammary epithelial cells. Diseases associated with ZnT4 include zinc-deficiency acrodermatitis enteropathica and acrodermatitis acuta. Pathways involved include metal ion SLC transporters. An important paralog of this gene is SLC30A2. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.6 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
Ohhh Mr. Iguana
Size: Medium/Large, Color: Illana the Iguana - Large
Mr. Iguana is a house favorite! I got my first one of these in a barkbox and my dog loved it so much, i went on the hunt for another one. Amazon saved the day! I own about 7 of these now. They are prefect for dogs just love to chew, they hold up great! And they are heavy duty, super easy to clean! My dogs are very happy, great value for the money! Doesn't have a smell, vibrant colors! Noise level is quiet, minus my dogs chewing !
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Reviewed in the United States on August 13, 2025
★★★★★ 3
Not for strong chewers
Size: Medium/Large, Color: Dog Ness Monster - Large
Initially, I really liked this for my dog. She is about 8 months old and still loves to chew and destroys most toys in a few minutes. When we got this, it seemed like it would hold up for a while despite being clearly gnawed on. Unfortunately, once she focused on the softer "water" portion of the toy, that is where things went downhill with this toy. After a few short chew sessions, she was tearing chunks out of it. We had purchased a replacement since she liked the original so much, and after about 5 minutes with the replacement, she had taken enough out of the blue section that we immediately threw that one out as well. Overall while the green portion held up, the blue didn't. I can't recommend this for strong chewers.
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Reviewed in the United States on December 8, 2025
★★★★★ 4
Definitely not indestructable
Size: Medium/Large, Color: Dog Ness Monster - Large, Size: Medium/Large, Color: Dog Ness Monster - Large
I have a 1-year-old toy fox terrier (Rocky, pictured). He has been chewing on this toy for seven months now; it is one of his favorites. He has been unable to seriously damage the "ocean" portion of this toy, but as you can see from the provided image, the sea serpent has not faired that well. It is missing its snout and smaller portions of the humps. My guess is that the advertiser images showing much larger dogs with "like new" versions of the toy are just that...NEW AND UNCHEWED.
I have not had to clean up bits of this toy from my floor, so I assume Rocky has been swallowing them. My hope is that the yellow material of the serpent is not chemically dangerous when exposed to digestive fluids.
Rocky easily destroys plush toys in a matter of days if not hours and, consumed pieces of a small microfiber blanket that he liked to toss around and play tug-of-war with (past tense as he destroyed it and has since been trashed after I began finding feces-stained undigested pieces of it lying around). Since this toy has survived seven months and has remained recognizable, I have to agree it is durable.
Just beware, if my small toy fox terrier can damage the toy as pictured, expect your large breed chewer to do similar or worse damage. Also, the toy is very heavy! When Rocky drops it down a flight of stairs, it makes a MAJOR thud at the bottom, and sometimes makes me wonder if it might crack the large tile at the bottom.
I cautiously recommend the product.
Note the yellow frisbee in Rocky's picture. It is only a few months old and surprisingly has not suffered any chew damage despite substantial abuse. It appears to bend in the wind like a proverbial reed, evading damage by yielding to pressure.
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Reviewed in the United States on July 5, 2024
★★★★★ 5
Best Option on the Market for True Power Chewers
Size: Large
My two Power Chewers never get through these! Every other “power chewer” toy doesn’t last more than a week, but Nylabone lasts for months. I don’t buy any other brand any more.
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Reviewed in the United States on June 5, 2026
★★★★★ 5
So worth it!
Size: X-Large
The dogs love them! They chew on them and carry them around inside and outside. They are very durable to the weather and a pit bull! Recommend
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Reviewed in the United States on April 9, 2026