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Description
Human twist1 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 twist-related protein 1 (twist1). 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 amount of twist-related protein 1 (twist1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Twist-related protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Twist-related protein 1 (TWIST1), also known as class A basic helix-loop-helix protein 38 (bHLHa38), is a basic helix-loop-helix transcription factor that shares similarity with another bHLH transcription factor, Dermo1 (also known as TWIST2). It is encoded by the TWIST1 gene. It is thought to regulate osteogenesis and is activated by multiple signaling pathways, including Akt, signal transducer and activator of transcription 3 (STAT3), mitogen-activated protein kinases, Ras, and Wnt signaling. Activated Twist can upregulate N-cadherin and downregulate E-cadherin. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
Based on 711 reviews
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Product Reviews
★★★★★ 5
Keeps ALL it's promises
Configuration: Vacuum
12/27/18 I have been using this vac over a year to clean our home's hardwood and vinyl floors and low pile area rugs. Disclaimer: we are not related to the company in any way, shape or form - have to say that because this review may sound like a commercial! This vacuum is small, quiet, very powerful and fulfills every promise the company makes about it. This unit comes with 3 small tools contained in the unit, 1 hard floor tool (vacuum powered) and 1 low pile carpet tool (vacuum powered). The 2 floor tools clip onto the sides of the canister. There are 6 power settings from a low for drapes to a high for hardwood floors, all marked with an icon. The power cord IS EXTRA LONG - yes, fewer plug changes - and the re-tractor works beautifully (I do let the cord slid through my fingers and catch the plug end to keep it from slamming into the back out of habit). The wand section is collapsible, which I just thought was nice for storage until I began using that feature while vacuuming in tight spaces, where you can change the wand length on the fly. The floor tools' articulated connection to the hose looks quirky until you begin using it; it makes the tool head extremely nimble in getting into tight spaces. The hard floor head has a skip tooth brush pattern around the edge which pulls in dust instead of pushing dust in front of it. The suction works all the way out to the ends - it will pull dust into the head from ALL the way around. The dust bag and separate HEPA filter have markers to let you know when to change them. I haven't changed either yet (note: we do not have pets, the house is vacuumed weekly). It tracks easily and the side bumper strips are placed where they actually work!
Problems? well it is expensive, but I don't see replacing it any time soon.
Seriously, I am still amazed at how well built and well thought out this unit is.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 27, 2018
★★★★★ 1
I absolutely hate this vacuum.
Configuration: Vacuum
I bought it due to the positive reviews and was tired of my Dyson ball vacuum. Since day one I have had nothing but problems with a vomit like smell that comes out of this vacuum every time we use it. It got to the point that we have replaced the filter with expensive carbon filters, washed out the hoses and attachments with bleach, and nothing has worked to get this thing to smell better. To make matters worse, the suction power of this thing is limited at best compared to other vacuums we have owned. I can go over the carpet with my old Dyson and pick up a significant amount of dirt that the miele cannot even come close to getting. I wish I had returned it when I still had a chance.
We have now bought a iRobot Roomba 980 and honestly it is 100x better than the Miele piece of junk that just sits in the closet now. It has strong suction power, is easier to clean, and keeps the house clean.
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Reviewed in the United States on January 21, 2019
★★★★★ 5
Nearly perfect!!!
Configuration: Vacuum
My Dyson ball cracked & I needed to replace. I despised my Dyson; it pushed more around than it picked up. I've been researching vacuums & landed on Miele but it took me a while to land on what kind. I have mainly hardwood floors & wanted something good to dust, do stairs & be a good total hope vaccuum.
I have allergies so C1 series out. C2 is too compact so I landed on C3 (great YouTube videos on all this).
This is a lower C3 end with the bells & whistles. I can click controls with my foot & I forgot how nice retractable cord is. What drive me to this model is the turbo brush head... this head had great reviews.
So I just got it & used & WOW. My floors feel so clean. The crevice & dusting attachments are great. I whipped through all my baseboard & trim in no time. It's light weight & doing stairs was easy compared to clumsy Dyson. The Jose is flexible & easy to work with.
The only con I can see, & I saw reviews on it & agree, there should be a handle to hold the vacuum head with v holding the curve of the hose. It feels ergonomically off when doing floors but it feels fine when dusting/stairs. Other con is that the hose doesn't stand up line an upright (which I expected since it's a canister) but that's really it.
The pros so far confirmed I made the right choice. I have a small dog but she doesn't shed. When selecting a model you need to think of what your needs are & what attachments come with each & go from there. Analise model has more features but I wanted an out of the box, w: right attachments ready to go.
It's an investment so I feel like I made the right choice on this one.
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Reviewed in the United States on November 28, 2017
★★★★★ 5
Great way to control TV and radio in one room from an easy chair in another room!
I help a 96 year old blinded vet with tech support -- and as his health worsens, needed to make his TV, his radio, and the audio switch that selects what audio should go into his hearing aids via Bluetooth work from his easy chair in his living room. All the equipment is in his bedroom. This IR repeater system installed quickly and easily and works perfectly. Now he can summon ball games, news, or whatever he wants without fear of getting up and falling down. A perfect solution!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
★★★★★ 5
Magic!!!
I am no tech expert but these little guys are magic! We had just put in a wall of built-ins and desperately wanted to hide our cable box behind a cabinet, but with it being infrared this would not work without these little repeaters! First off I just have to say that the Sewell customer service is off the charts. You actually get to speak to a real person right away, no annoying menu after menu trying to get help. They helped us so much when we hit a couple snags. Here are a couple things that I figured out that I hope will help others. First you can hide both of these little discs, they both come with tiny little eyes that can attach to your tv and cable box or whatever you are trying to use them for, and they are nearly invisible. Second, don’t get nervous when you plug them into the power outlet and the lights don’t come on, this is ok and they aren’t supposed to stay on, they kind of blink here and there, and only when you use your remote. Third, we have an Altice One cable box (not the mini, just a regular cable box) and it was impossible to find the eye to attach the emitter’s eye (Side note: we had to put the emitter disc on top of the cable box because we couldn’t fit the emitter disc pointing in front of cable box. The directions give you both options though and you can basically put it anywhere as long as the eyes attach). The phone trick did not work on this particular cable box. But after trying about a dozen different spots on the front of the box we finally got it to work by putting the eye right on where the digital clock numbers are. I put it basically right on top of the colon (:). Cable vision was no help with this so I hope this can save others from our frustration. After we found the eye everything came together and worked perfectly! No more do we need to keep the cabinet open to change channels… it’s a miracle!!! Hope these little guys work forever.
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Reviewed in the United States on November 22, 2022
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