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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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4.1 ★★★★★
Based on 2021 reviews
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Product Reviews
★★★★★ 5
One of the best
Scent: Bourbon Oak
One of my go to and you won't regret it
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 4, 2026
★★★★★ 5
An incredible guide for your Advent journey.
Format: Paperback
Thank you Miriam for bringing us this beautiful and deep guide for our Advent journey. I used it for the first time in 2023, and will definitely be using it again in 2024. I loved and looked forward to every day that I sat with the daily passage.
Miriam guides us into daily Scripture, and includes questions for us to meditate on and apply those passages. In addition, she adds her own meditation, which offers a rich perspective. It is accompanied by Lilias Trotter's own perspective through her words and wisdom and gorgeous art. The author invites us into self-reflection multiple times through each daily passage and includes poetry and song that binds each daily message.
I will definitely give these as gifts for next year, and I am so looking forward to walking through it again next Advent. It will be interesting to see the Lord's living and active wisdom reveal itself year after year.
I would highly recommend keeping a journal as you dive in to this rich, rewarding journey.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 27, 2023
★★★★★ 3
Good news, bad news.
Format: Paperback
I like the product, but because it was inadequately packaged, the top of the book is imperfect. It's certainly readable, but if I am paying full price, I expect a perfect copy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 24, 2024
★★★★★ 5
Deep Into the Financial Nuts and Bolts
Format: Paperback
This a very, very informative book about the financial crash of 2008 and its lengthy recovery. The British author is a fine writer and definitely appears to master the subject. His background is as an economic historian and his approach to the financial collapse is thru the lens of political economy.
For a European, he is very charitable towards the failures of the US markets and the responses of the US govt in the crisis. In fact, he is much more critical of the Germans and the British and the EU govts in their involvement in the crisis and their failure to more effectively guide the Eurozone out of the financial crisis.
The book is somewhat dense and daunting and is 616 pages of text. Definitly not for the faint of heart or those who are not somewhat financially and economically literate. It does not lend itself to speed reading. A glossary and list of abbreviations would have been very helpful--as other commenters have suggested. Abbreviated acronyms of organizations, treaties, agreements,govt agencies, etc. come flying fast and furious. Still, with due and diligent effort on the part of the reader, you can see how very well the author knits his technocratic financial narrative together.
The negative reviews often seem poorly informed. It's a difficult book to get thru. Some readers appear to dislike the author because he is a Keynesian liberal. I understand that, but they should note that he tends to be very positive about US govt financial leadership leading the world out of the crisis. Some don't like the fact that he thinks Trump is a grifter. Unfortunately, Trump is, irrefutably, a grifter and a con man.
The only areas where I found myself disagreeing with the author were on the issues of immigration and protectionism. The Eurozone and the US cannot support unrestricted immigration. There are too many millions in the world of 8 billion people who would kick down the gateways of either entitity to gain entry. Unrestricted immigration is an existential threat to all the Western countries involved. Sober assessment of reality is something other than "racist". "Realistic" is a far more accurate term.
The author is a classic liberal and against national "protectionism". However, this is 2023. The book was written in 2017. Much water has passed under the bridge since then. Countries are political entities with borders. Too often--as the writer points out--economic and financial advisors forget the "political" part of "political economy". Trump's legions are there to remind them. At least they are good for something. Protecting your economy from essential foreign supply chain failures and from hostile foreign competitors is just good governance. And insuring that others share in general prosperity besides coastal elites is another part of good governance. "Flyover Country" is every bit as important as the two coasts. Probably more so. Never forget the "political" part of "political economy".
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Reviewed in the United States on July 29, 2023
★★★★★ 5
A Complete Autopsy of the 2008 Financial Crash and the Government Response
Format: Hardcover
First, let me say that I can't believe one person wrote this entire book. Tooze handles everything - economics, finance, politics, diplomacy, public policy, housing, discrimination, trading platforms - and does it expertly. The author's detailed understanding of such a wide arrange of topics is dazzling, especially his handle on inscrutable national bank mechanics.
Second, if you want a macro understanding of how governments responded to the 2008 financial crisis and how these responses produced such wildly differing results, read this book. The analysis at times is like eating sawdust and it is excruciatingly detailed. There were entire chapters on the European side of the crises that I felt were repetitive and could possibly be removed from the book without losing too much. However, clearly Tooze has done his homework and the data underlying his conclusions is vast.
Third, I learned a lot and I had previously read "Too Big to Fail" by Sorkin and some other lesser known books on the recession. Tooze handles everything from a policy perspective and his data support his overarching theme: the US had a cohesive, massive stimulus program that probably could have gone further, while the EU responded in nibbles. The US rebounded well, albeit not perfectly while the EU went from crises to crises. He also shows that there were massive consequences from the bailout of the financial system: Brexit and cartoonish clown politicians like Trump in the US, and Farage in the EU, who are supported at their roots by a dangerously racist and nationalistic surge of those left behind by the modern global economy. This is an excellent read.
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Reviewed in the United States on November 10, 2018