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Description
Mouse GLUT4 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 glucose transporter 4 (GLUT4) capture antibody. 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 glucose transporter 4 (GLUT4) 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 Glucose Transporter 4 | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glucose transporter 4 (GLUT4), also known as solute carrier family 2, is a protein encoded by the SLC2A4 gene. It is an insulin-regulated glucose transporter primarily found in adipose tissue and striated muscle (skeletal and cardiac muscle). The gene encoding GLUT4 was cloned and mapped in 1989. On the cell surface, GLUT4 allows circulating glucose to diffuse along its concentration gradient into muscle and adipocytes. Once inside the cell, glucose is rapidly phosphorylated by glucokinase in the liver and hexokinase in other tissues to form glucose-6-phosphate, which then undergoes glycolysis or is polymerized into glycogen. Glucose-6-phosphate cannot diffuse out of the cell, maintaining the concentration gradient that allows glucose to passively enter the cell. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatants, and other biological fluids |
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4.5 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
All 3 Dogs Want The Benebone Ring Now
Flavor Name: REAL Bacon, Size: Large
Rescued an Old English Sheepdog and after about 2 weeks, he started chewing things in the house. We knew he enjoyed chewing sticks in the backyard, but chewing things in the house was a no go. Purchased this Benebone Ring (bacon) hoping it would help to curb some of the issues we were having. I am happy to report that so far, after 3-4 weeks of him having the Benebone Ring, he has not chewed a single thing inside the house. He loves this ring and carries it in and out during the day so that he has it with him. The only problem we ran into was our other 2 dogs ALSO wanted the Benebone Ring. While they are not big chewers, suddenly this toy became the must have item in the house. So of course, we had to get more.
Highly recommend this chew toy as it is not only durable, it appears to be highly desirable. Keep in mind however that this toy is heavy. So when your dog decides to drop it on your hardwood floors, you WILL hear it in every room in the house!!
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Reviewed in the United States on March 18, 2026
★★★★★ 5
My babies love their Benebones!
Flavor Name: REAL Bacon, Size: Large, Flavor Name: REAL Bacon, Size: Large
My one-year old pittie is 60lbs and has severe allergies to EVERYTHING, she's on medicated food and all the crazy things, and so I was trying to find something that wouldn't mess up her regimen by her ingesting it. This is PERFECT. She loves her medium sized Benebone! She carries it everywhere haha. I have three other big dogs, and got the largest one for my 110lb German Shepherd and she also loves it. I keep an eye on the edges and sand them down if I find anything sharp.
These things are TOUGH. My pitbull can eat a massive bully stick in less than ten minutes, and she's barely made a dent in this thing! My GSD is a little more delicate and likes to savor and enjoy her toys, so hers isn't progressing as fast. She eats Yak Chews in around 30 minutes (and those things are like concrete!)
I think these will easily last a month, possibly more, and for the price you can't beat it. The value is insane. I spend maybe 200$ a month on rawhide alternatives and bully sticks and yak chews for them, and these 14$ things last infinitely longer. BTW these things hurt like a mother trucker when you step on them, so keep an eye on where they are haha they're big and heavy!
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Reviewed in the United States on November 9, 2025
★★★★★ 5
Love these for super chewers
Flavor Name: REAL Bacon, Size: Large
I have two pit bulls and they destroy everything. These have held up very well for weeks. I use the cups for their morning yogurt and can even use hot water wand soap to wash them out. These are a great value for the money and the dogs love to carry them around in their mouths and even tug o war with them. Highly recommend.
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Reviewed in the United States on April 15, 2026
★★★★★ 3
Good
Flavor Name: REAL Bacon, Size: Medium
Decent product, my dog likes it, except I think it’s made of some kind of plastic
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Reviewed in the United States on March 3, 2026
★★★★★ 5
Amazing
Flavor Name: Peanut Butter, Size: 6.9 Ounce (Pack of 1)
My dogs love these so much, seems to keep my girl meatballs teeth clean. Both of them go craft for these, they chew on them almost all day, it keeps them busy. I bought other brands of these but they had this jelly filling that wasn’t safe for them, so I switched to this peanut butter one that seems a lot healthier and is a lot safer for my dogs (in a sense they won’t choke on it from basically inhaling it again 😭) and they are still interested in the bone once it’s empty, so it’s a toy that lasts forever. Overall an amazing cheap dog find on here
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Reviewed in the United States on June 7, 2026