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Description
Human SLC39A4 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 ZIP4 (SLC39A4). 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 zinc transporter ZIP4 (SLC39A4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Zinc transporter ZIP4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Solute carrier family 39, member 4 (SLC39A4), also known as zinc transporter ZIP4 (ZIP4), is encoded by the SLC39A4 gene. It is a transmembrane protein that has been associated with acrodermatitis enteropathica. As a zinc transporter, it plays an important role in cellular zinc homeostasis. This gene encodes a member of the zinc/iron regulated transporter-like protein (ZIP) family. The encoded protein is localized to the cell membrane and is required for intestinal zinc uptake. Mutations in this gene cause acrodermatitis enteropathica. Multiple transcript variants encoding different isoforms have been identified for this gene. | |||||||||||||||||||||||||||||||||
| 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 600 reviews
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Product Reviews
★★★★★ 1
These are cheaply made, and garbage, highly dangerous letting a dog chew on a battery pack
Color: Grid Pattern
These units are garbage. I got one for each of my dogs charged it up turned it on put it inside the rubberized ball shell and securely fastened it less than two minutes later my dog had taken it apart by just biting it and then started to chew on the battery and motor unit. So if you wanna have a lithium polymer fire in your house and potentially kill your animal, go ahead and get this, but if you do not want a fire in your house and you want your animal to live then absolutely do not buy this garbage it’s cheap we made and it’s not strong enough for any dog
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 27, 2026
★★★★★ 4
Good Product, but images over promise just a little.
Color: Ripple Pattern
- Performs well on tile and low pile carpet. On artificial grass it doesn’t roll even on the highest setting; it only vibrates. Watch the video.
- Still fun for my pup. I’ve only had it a day so no clue about durability although my dog is a strong chewer.
- Price Value: When purchased it was 2 balls for the same price of a single ball from competitor.
- Material feels like a Nerf football. Easy to disassemble and charge.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 16, 2026
★★★★★ 5
My dog enjoys it
Color: Ripple Pattern
My dog loves the moving ball! She go chase around it and pretty durable. Comes with two balls in one set.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 27, 2026
★★★★★ 5
Best jiggle balls we have found thus far
Color: Ripple Pattern
Yes ! First off this was a great value for the money. The balls function just as described and they have a decent battery life. They entertain both our goldens for a good bit of time. They have so far held up and not been chewed up . We limit the time and put them away after that time.. to avoid them being destroyed. Of all the jiggle balls we have purchased these are by far the best. The way the component is encased in the rubber protects it.. and it screws together very well. I am so pleased with these !
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Reviewed in the United States on December 29, 2025
★★★★★ 5
Love this set!!!
Size: Medium (2.5"), Style: Fetch Pack 1
I was trying to figure out which of the many different balls this company offers, and I was getting frustrated but then i was SOOOO happy to see they had multiple sets that had multiple of the different balls they offer!! I bought this set and the other set they offer, but this one was my favorite and I think my dogs too! The orange ball works great and bounces just high enough that it doesn’t clear our patio wall, but enough for him to go and jump up to retrieve. But the glow in the dark ball is his favorite! Both me and my pomsky were blown away how we brought the glow in the dark ball outside just as the sun was going down, and when we brought it back inside it was BRIGHT AND GLOWY!!! Even with the sun going down and not much light left!! He loves to play with it in the house after it’s all glowy from the sun. Sometimes we even glow it up under a light inside the house if it’s dark outside, and he will play with it outside in the dark haha he loves it!!! And I think it’s pretty cool too!! All of the balls this company overs are VERY durable! We realized early on that we cannot give our dog regular tennis balls cuz his favorite thing to do is tear the outside fabric off!! So we were trying to figure out what company has the best balls for dogs so they can’t sit there and try to pry them apart, and we found this company!! We have already bought from them about 3 times and all of their balls are durable and bounce how they say they will bounce!! They have the super bouncers and the glow in the dark one and the one with angles that will bounce in different directions and the ones you can throw REALLYY far! Overall, all the balls from this company are very durable, bounce really good, and keep my dog occupied without allowing him to sit there and tear it apart!!! Which i love cuz I do not like when he stops playing with us to sit there and try to take off the fabric, cuz then he could swallow it! So always buy the balls from this company!!! They are the best and most durable!!!! Also plenty soft for your dogs mouth so they can chew on it a bit and it won’t hurt their teeth/gums!
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Reviewed in the United States on July 28, 2024
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