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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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4.2 ★★★★★
Based on 738 reviews
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Product Reviews
★★★★★ 5
Angels and the Fallen — Breathtaking Start
Format: Kindle
This book was EVERYTHING I expected it to be and more. Leave to R.L. Caulder and M. Sinclair to give us yet another amazing book!
Kieran and her guys hooked me from the very beginning and did not let go. Found family is one of my favorite tropes and it was done so well in this book. Couple that along with an FMC trying to find her place in the world and I was practically drooling over this book. The world building, the conflict, the wyverin sidekick — it all was done so well that it felt fresh and real.
Each love interest felt real and unique in their own ways. I felt connected to each one and like they were truly different people. I love that so much in a book and these two authors never miss with their love interests.
Multi POV, Reverse harem, who did this to you, and a magical world highlighting the angels and the fallen. This book has everything and it does it so well.
Big thank you to R.L. Caulder and M. Sinclair for the arc copy! I cannot wait for book 2!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 20, 2024
★★★★★ 4
A new take 🤔
Format: Kindle
I’m not going to go into what this book is about — by now you e already read the synopsis and have an idea — but I will give a couple of insights.
Definitely a different take on angels and their fall; I like it. This is an interesting start to a new series - YES, this does end on a cliffhanger so be prepared for that. I’m eager for the next book (and hopefully the last in this series — sometimes the author(s) will switch it up depending on how the story flows for them) in this series.
I’m hoping that a certain male character redeems himself because he’s a dick that will make the meaning of mixed signals jealous lol.
You ever watch that old movie — I think it’s call Mommy Dearest? — yea, let’s switch that up to daddy. That man would make a perfect husband to mommy dearest.
I recommend this book because while these authors are well known for their cliffhangers, they are also well known for putting out good stories.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 19, 2024
★★★★★ 3
Liked it
Format: Kindle
I liked the worlds and the characters. The prophesy is confusing in that I would think, since all worlds ore on a path to destruction, the angels would want their world to be saved. I am sure there is more evil that will be revealed as the series proceeds. I dislike a prolonged “poor little me” trope and the main character has moved past that, thankfully, and began to show a fierce attitude as the novel progressed. The last portion of the novel had some great action. It’s worth reading, just be patient with a slow start.
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Reviewed in the United States on February 14, 2024
★★★★★ 5
Could not put this book down!
Format: Paperback
Binge read quality!! Finished in 2 days I was in a dilemma with this book. I couldn't wait to finish because it was so good, and yet I didn't want it to end, because it was so good. Staying up til 2:30am is the highest complement I can give to a book. Current day with Gothic feel. The story pulls you in immediately, and there is never a part where it feels dull, or that you need to skip through. Mysterious with plot twists, especially the end. I love how there were flashbacks from the past intertwined with present day. Gave a great backstory that only came together at the very end. Spicy for sure, and fit in perfectly with story. Great character development so that you could empathize with both of the main characters.. Lucian and Isa. Check Trigger warnings first!
Highly recommend this book
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Reviewed in the United States on March 10, 2026
★★★★★ 4
Easy Read With a Wild Ending
Format: Audiobook
This was a really easy book to get pulled into. The writing flowed well and I flew through it without feeling bogged down at any point. While I don’t think it hit me as hard as some of Keri Lake’s other books, I was still invested the whole time and needed to know where everything was going.
The atmosphere and mystery kept me hooked, but the ending? That twist completely caught me off guard. I honestly did not see that coming at all. Even though this one wasn’t my favorite from the author, the final turn alone made it worth the read.
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Reviewed in the United States on May 16, 2026
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