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Description
Human GSK3b 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 Glycogen Synthase Kinase 3 Beta (GSK3b). 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 Glycogen Synthase Kinase 3 Beta (GSK3b) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Glycogen Synthase Kinase 3 Beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glycogen synthase kinase 3β (GSK3β), encoded by the GSK3β gene, is an evolutionarily conserved serine/threonine kinase ubiquitously present in mammalian eukaryotic cells. In addition to its earliest known role in regulating glycogen synthase (GS) activity, it also acts on numerous signaling proteins, structural proteins, and transcription factors to regulate cell differentiation, proliferation, survival, and apoptosis. It has been selected as a therapeutic target in the research of a variety of major diseases, including cancer, neurodegenerative diseases, and neuropsychiatric disorders. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.9 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 3
Weird smell
Color: White-cooling Basic, Size: Queen(Pack of 2)
The pillows are great for someone looking for something cool and comfortable. We loved them so much that we bought a second pair. However, we were seriously disappointed with the odor of our last purchase. They give off a strong and indescribable chemical/burning plastic smell so nauseating that it’s difficult to sleep with. We’re hoping this issue will resolve with a couple more washes of the pillow cases, but am worried the source is the stuffing itself.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
helped treat my sore neck
Color: White-cooling Basic, Size: Camping Pillow(Pack of 1)
This pillow — after dialing in the right amount of shredded foam — has greatly helped my sore neck. Initially, I had to remove some foam. Then I gradually added foam until it reached optimum comfort and support.
It is also CPAP-friendly — especially when I position my head near the pillow's edge. (I am a left-side sleeper.)
I like the travel size because I use a bulky hugging pillow to ease stress on my shoulder and elbow. That makes my side of the bed feel cramped if using a larger head pillow. A smaller pillow is easy to flip end-to-end in the middle of the night.
I have spent lots of money on pillows over the years, but this is the best I've ever had for relieving [1] neck pain and [2] CPAP mask seal problems. Very comfortable, supportive, and affordable.
I did not notice any unpleasant smell — and I am usually sensitive to that kind of thing.
Would highly recommend.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Love them!
Color: White-cooling Basic, Size: Queen(Pack of 2)
Love them! After 10 years with our MyPillows, they finally lost their fluff. The reviews I read of the MyPillows of today were not good; they didn’t have as much filling as they used to. These pillows remind me of how fluffy our new MyPillows were 10 years ago!! You sink right into them and can mush them however and they stay. These are way cheaper than the Coop and it’s the same concept; add or remove the filling as needed. The cooling side is a huge plus too! You won’t be disappointed with these pillows!
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Soft and cool.
Color: Cooling Blue Cover, Size: Queen (Pack of 2)
Love this pillow set. Comfortable addition and nice and cool at all times. Going to be great for when the hot weather hits
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Reviewed in the United States on June 8, 2026
★★★★★ 5
As advertised for once! Lol
Color: Cooling Blue Cover, Size: Queen (Pack of 2), Color: Cooling Blue Cover, Size: Queen (Pack of 2)
Queen size set of 2 pillows, I'm giving it 5stars first bc it has a good length and width!! If you've ever bought memory foam pillows before you know some are small!! Anyways after having it for 5nights, we loved that me and my huddy could adjust the Thickness amount we had in it to our needs, me needing a thick pillow and my husband needing a flatter one. So this was perfect for that. So far its nice to sleep on, stays cool, doesnt get hot. I would say its on the frimer side but softer the more foam you take out! Really its just good for the amount you spend!! Will it last 10years probably not but what does now a days. This is the only time my huddy n I bought the same pillow and both love it so I'm just still in shock it worked out so well!!
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Reviewed in the United States on April 28, 2026