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Description
Human FOXO1 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 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 Forkhead Box Protein O1 (FOXO1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Forkhead Box Protein O1 (FOXO1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Forkhead Box Protein O1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Forkhead box protein O1 (FOXO1), also known as forkhead in rhabdomyosarcoma (FKHR), is a protein encoded by the FOXO1 gene. It is a transcription factor that plays a crucial role in the regulation of glucose metabolism and glycogenolysis by insulin signaling and is central to determining whether preadipocytes commit to adipogenesis. It is primarily regulated through phosphorylation at multiple residues, and its transcriptional activity depends on its phosphorylation state. FOXO1 may play a crucial role in apoptosis, as it is phosphorylated and inhibited by AKT. Furthermore, FOXO1 transactivates Bim, a member of the Bcl-2 family that promotes apoptosis and plays a role in the intrinsic mitochondrial apoptotic pathway. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.0 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Works just like it says.
Color: 8-in-1 Silver, Size: Rechargeable
My hands can no longer open a bottle of wine. This makes it so easy.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Great device
This wine opener is great. Is small and feels solid It really works. I was also impressed by the automatic dumping of the cork.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Best electric corkscrew - by far
I have tried others but this electric corkscrew outshines them all....so far. Really like it. I have been disappointed with some before though if you get a very dry cork that splinters.....but this one has a much smoother operation and just works better. Good price too.
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Reviewed in the United States on April 14, 2026
★★★★★ 5
It is light weight, rechargeable and it works hands-free.
This wine bottle opener is well designed and well constructed. It comes with a foil cutter and works hands-free when you push the button. It pulls up the cork - and then pushes it back out to discard the cork. It is rechargeable!! It is replacing an older opener that left behind cork fragments in the wine - and ate AA batteries quickly.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
Best on the market!
I have tried three other brands of electric cork pullers. The fact that I now invested in a fourth probably telegraphs what this review is all about: this Vinabon device is great, hands down winner in the electric cork puller sweepstakes!
First, its motor is more powerful. Some winemakers are employing corks that fit pretty tightly, and the other brands I used would occasionally be unable to remove the cork. The screw would simply stop turning, without ay way of resuming. Advantage: the bottle.
Second, it is small, about half the size of the others, leaving a small footprint on the counter.
Third, it lasts much longer on a single charge. It doesn't sit in a cradle, because it doesn't need to. Assuming you open 10 to 20 bottles a week, you will be able to go two weeks without needing to plug the charging cable in, and a full charge does not take long. 4 vertical lights on the cylinder tell you how much of a charge remains.
Fourth, you simply press and release the start button and maintain gentle pressure on the bottle while the device descends into the cork and removes it. As soon as you remove the device from the bottle, it immediately begins to eject the cork. All with that one touch of the button!
Fifth, if you encounter a particularly tight cork, the device will stop, whereupon you push the button again and it continues. A couple of synthetic corks required me to restart 3 for 4 times, but it ultimately conquered them all!
That leaves only one issue: a failed cork that breaks during the removal process, because the screw cannot engage the cork enough to remove it without it breaking. To my knowledge, the only solutions to this, with any cork puller, are to resort to an Ah So, or if that cannot do it, just shove the cork down into the bottle, and drink the entire bottle in one evening.
Truth to tell, I have, myself, actually enjoyed that latter remedial step, especially with a larger format bottle on an evening when I am dining alone.
I cannot yet report on the ultimate problem that seems to vex all electric cork pullers I have owned: If the cork disintegrates within the cylinder during removal, how do you get the remains out? I threw my last cork puller away because I could not remove the cork detritus stuck in the cylinder, rendering the device useless thereafter.
If there is a solution to that, I would not be surprised to find that this unit already handles the problem.
If BMW were to ever make an electric cork puller, I expect it would be much like this one.
t costs almost twice as much as the others I have owned, but you get what you pay for. After all, you can buy a small fleet of Yugo's for the price of a BMW.
Bravo, Vinabon! I
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Reviewed in the United States on September 17, 2024