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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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4.8 ★★★★★
Based on 2138 reviews
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Product Reviews
★★★★★ 4
Romance and Family Drama
Format: Kindle
Julia finally returns home after years away because her mom is dying. Going back to Gull’s Harbor, California means facing a lot of memories Julia would rather leave behind, including her high school sweetheart. After their mom’s passing, Julia and her sisters are in for a rude awakening when the will is read. The girls must live together in their family home for the three months or they forfeit their inheritance. Living in her childhood home puts Julia right next door to her high school boyfriend who she’s still in love with. Will Julia be able to win Liam back or are there too many secrets between them?
I have only ever read cozy mysteries by Jenn McKinlay, but was intrigued when I saw this book coming out. I loved the cover and couldn’t wait to dive in. I liked Julia and Liam together and enjoyed the second chance at love for them. The will and storyline that brought on led to some drama and funny moments. This was a romance book and had a lot of good moments between Julia and Liam, but it was also a story about finding yourself after grief and family. I liked the different layers in this book and it kept me interested from start to finish. The ending of this book was great and I loved how everything tied together. The family dynamic between the three sisters was fun to read about and I loved seeing how all of their lives were so different even though they were raised by the same woman. The romance parts were the best in my opinion, but I did enjoy the variety in this book and had a good time reading it.
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Reviewed in the United States on July 17, 2025
★★★★★ 5
Nice read about familial relationships, grief, and romance.
Loved this book - quick, cute, easy read. Stories about handling grief can sometimes be overwhelming or triggering but this was well done and speaks to all ranges of mother daughter relationships.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 5, 2025
★★★★★ 5
I loved this book!
Format: Paperback
I really enjoyed this well written book, “I Can’t Even”. It covers family topics of love, loss, heartbreak and diversity. There are some laugh out loud scenes and plots that will keep you guessing right until the end. I loved it!
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Reviewed in the United States on May 3, 2025
★★★★★ 3
Second chance romance
Format: Audiobook
I have loved every single book that Jenn McKinlay has written - so I was delighted to get a chance to read this one early.
Julia returns to her childhood home with her two sisters, when her mother Babs gets sick. Julia and her Mom have been almost estranged in recent years, and her mother doesn't seem very glad to have her home. When her mom passes away, the terms of her will reveal some family secrets, and some strings attached to the sisters and how they need to live, before they can inherit moms home and money. Cousin Paisley is hanging in the wings hoping to grab what the sisters should be getting.
While home, Julia tries to rekindle her romance with boy-next-door Liam, the guy she had been dating when she upped and left without a word many years ago.
We follow Julia and her sisters as they rebuild their relationships, and grieve the loss of their mother.
I liked the premise, and really hoped to like the book. Unfortunately this one just wasn't for me. Julia and Liam are just nasty to each other, and most of their issues could be solved with some simple communication. The descriptions of some of the other characters bordered on the mean rather than funny snarky. It seemed that most of the characters were using each other.
I listened to the audiobook and have nothing but praise for the narrator, who handled all the different voices really well and kept me interested in following the plot.
I hope this was just a blip from this author - as I am really looking forward to her next novel!
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Reviewed in the United States on July 12, 2025
★★★★★ 5
Glowing Review
Color: 8 Oz Browning Lotion w/ Body Tanning Brush
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Within just one use, I noticed a rich, natural-looking tan developing. It builds beautifully and doesn’t leave behind any sticky residue. Whether you're prepping for a beach trip or just want that sun-kissed goddess glow year-round, Maui Tanning Lotion is IT.
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⭐️⭐️⭐️⭐️⭐️ 10/10 – Take me back to Maui... every time I use this product!
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Reviewed in the United States on July 2, 2025