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Description
Mouse TRPC 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 classical transient receptor potential (TRPC) capture antibody. After incubation and washing, the color is developed using the substrate TMB. TMB is converted to blue by HRP catalysis and to yellow by acid. The intensity of the color is positively correlated with the classical transient receptor potential (TRPC) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse classical transient receptor potential ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | TRPC channels (TRPC) are a group of ion channels that are primarily located on the plasma membrane of animal cell types. Most of these channels are classified into two main groups. The first group includes TRPC ("C" for canonical), TRPV ("V" for vanilloid), TRPVL ("VL" for vanilloid), TRPM ("M" for mechanoreceptor), TRPS ("S" for solomelatin), TRPN ("N" for mechanoreceptor potential C), and TRPA ("A" for ankylosing). Group 2 includes TRPP ("P" for polyvesicular) and TRPML ("ML" for mucin). In vivo, some TRP channels are thought to behave like miniature thermometers that are used in animals to sense heat and cold. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.5 ★★★★★
Based on 602 reviews
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Product Reviews
★★★★★ 4
These are very thick pillows.
Item Package Quantity: 2, Size: Queen
Exactly as advertised. They are very firm and very thick. They're too thick for me to be able to use as bed pillows. However, they seem to work pretty well as cushions in my papasan chair on top of its normal cushion also work very well as decorative pillows.
I'm still giving them four stars because they're exactly as advertised and they seem well made. My only complaint is that they're just too thick. And that's definitely not the fault of the pillows or the pillow maker.
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Comfortable, firm, great for positional sleep therapy
Color: Cool Grey, Size: 24x24x12 inch
This is just what I needed to breathe better in my sleep, my doctor recommended a pillow like this for sleep apnea symptoms. I have an upcoming surgery too, so i'm glad I was able to test it out a week prior and see how it can help me prop up in bed. Feeling better about my upcoming recovery knowing i'll have this great pillow to support me!
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Literally perfect and exactly what I needed!
Color: Cool Grey, Size: 24x24x12 inch
I can't recommend this enough! It was exactly what i was looking for! I have an annoying habit of researching everything for months before I purchase something that will get used this often. I finally decided on this one and I was very pleasantly surprised. I needed a wedge pillow to put at my back while sitting in bed. Mostly for when I'm working on my laptop. This one was the perfect height and perfect fit for me.I'm a female that is 5'4 and weigh about 117pds. I wanted something firm enough to support my back, and this most certainly does! It had a very pleasant appearance with the light gray sides. It has a removable cover I can take off and wash when needed. It's portable enough to move from room to room, also works great on my couch and it has amazing versatility. It can also be used as a pillow for someone that might need to sleep at a slight incline. It also works if you need to prop your legs up.
It came vacuum packed. Once opened It said to let is sit for at least 24-48 hrs to give it time to properly inflate. For me, It plumped right up immediately! Again, from someone that does her research, if your looking for something like this at a reasonable price, this is it!
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Reviewed in the United States on May 10, 2026
★★★★★ 5
It was worth every cent
Color: Cool Grey, Size: 24x24x12 inch
I have BPPV. It's a condition where tiny ear crystals come out of place and move within your ear canals - this fools your brain into thinking you're in motion, causing severe dizziness. The most common treatment is the Epley Maneuver, and that actually helps a lot. However, I was going through a period where every single day I'd wake up with severe dizziness. Even if I managed to do the Epley Maneuver and it worked, the entire rest of the day would be ruined because I'd still feel generally unwell and nauseous. I ended up getting a book about BPPV and the author strongly recommended sleeping at at least a 30 degree angle. So, that's why I ended up buying this wedge pillow...
I started using it April 26 and today is May 17. Since I began using it as a pillow, I have not woke up dizzy again. That is so huge! It may be that sleeping at a 30 degree angle deserves 5 Stars but thought I'd give this pillow 5 Stars too, as it's been a great experience. If you suffer from this, here's how the angle should be...If you lay your head on a pillow and open your eyes, you should not be looking at the ceiling. With the right angle, you should be looking at where the wall meets the ceiling. If so, you know you're at the right angle.
As far as pillow itself, I'd say the pillow case fabric is of average quality. Not really remarkable in any way. It seems decent. Good value for the price. The inside is firm (which is just what I needed) but comfortable, and I tend to not move around much the entire night. I like this very much and if it does start wearing out, I'll definitely come back here and buy another. Would recommend.
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Reviewed in the United States on May 17, 2023
★★★★★ 4
A bit too firm but great otherwise
Color: Cool Grey, Size: 24x24x12 inch
I got this wedge pillow primarily to have something to lean my head against (I like to lay on my side when reading or looking at my phone at night sometimes), or to sit upright when reading in bed. It's very comfortable doing both now. It does a really good job at being a nice support for the back of my bed. Only negative is I feel it's too firm. I wouldn't use it as a primary pillow, but it's definitely good for back support at least.
While I won't mark it down for this, I do find it very large. If you have anything smaller then a queen it may take up more of your bed then you'd like.
Definitely recommend. Hopefully will last quite a long while as well.
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Reviewed in the United States on April 11, 2025
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