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Description
Rat CHRNa1 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Cholinergic Receptor, Nicotinic, Alpha 1 (CHRNa1). 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 Cholinergic Receptor, Nicotinic, Alpha 1 (CHRNa1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cholinergic Receptor, Nicotinic, Alpha 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Muscle nicotinic cholinergic receptor α1, also known as nAChRα1, is a protein encoded by the CHRNA1 gene. The protein encoded by this gene is a subunit of certain nicotinic acetylcholine receptors (nAchRs). Muscle nicotinic cholinergic receptors are composed of five subunits of four different types: two α isoforms and one each of β, γ, and δ subunits. The α subunit encoded by this gene plays a role in acetylcholine binding/channel gating. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.4 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
Dries completely clear, no pilling/balling, does have fragrance
Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen, Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen
I use a ton of sunscreen, religiously, and this one, at SPF 50, caught my eye. I never heard of this brand before or tried any of their products. While I am not a kid, I often like to use kids' sunscreen because I find it to be more gentle. This is a mineral sunscreen with zinc oxide as the filter. Mineral sunscreens often leave a white cast but this one claims to dry completely clear - and I was curious to try this. Indeed, it dries completely clear! It absorbs amazingly and is a beautiful sunscreen for the face. It leaves a nice velvety glow when it dries - there is no oiliness and no shine. Even if you wear no makeup or products on top of it, you will look perfectly fine. I also tried layering it with multiple products, both under and over, and this sunscreen does not pill - which is another huge plus for me - as there is nothing worse than starting over when your sunscreen makes your face look like an odd mess. Overall, the performance of this sunscreen completely blew me away - this is one of the best sunscreens I've tried. A word of caution though - this sunscreen does have fragrance (and it is pretty heavily fragranced) so do keep this in mind if you are sensitive to fragrance in skincare.
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Reviewed in the United States on February 12, 2024
★★★★★ 5
50 SPF Aloe Up Suntan Lotion -- 3 ounce Tube -- Smells great! :-)
Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen, Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen
Sunscreen is super important because it allows you to stay in the rays of the sun. This tube of 50 SPF Aloe Up has been great so far! The tube, at 3 ounces, is a prefect size to keep in a toiletries bag. There was a seal under the cap and the tube has a small hole, which helps prevent unintended over-squeeze waste.
The lotion is a bit thinner than I expected, but it rubs in smooth/invisible and smells great! I had no problem with breakouts or clogging of pores, and no burn under winter sun. With regard to winter sun, I rated this a 5 since so far this has worked great and there's been no problems...But... because we're under winter sun right now, I won't be able to test this under full strength sweaty summer sun for some time. I will come back and update this review if anything changes in the heat and stronger sun of summer. Hope this helps! :-)
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Reviewed in the United States on February 27, 2024
★★★★★ 5
Quality sunscreen with a light scent and no griminess!
Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen
Most sunscreens I have used in the past have a very distinct smell and/or leaves excess oiliness residue, so you can imagine I have my doubts when a manufacturer advertises its product have a clear and lightweight application with hydrating and comfortable feel on application. To my surprise, this product actually delivers on its claims! The consistency of the lotion itself is indeed very lightweight and easily works into the skin without being greasy as normally found with other brands. Despite its light application, I found that it still offers great protection after being exposed to moisture or coming in contact with water. A sunscreen with a relatively high SPF protection in a non-greasy, lightly scented and weighted application along with its accommodative formulary for sensitive skin is quite impressive; and about the only negative I can find is its high price for a 3 oz. tube relative to other brands. Overall, this is a good sunscreen at its list price, and one that I can recommend if you can find it at a discount.
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Reviewed in the United States on March 13, 2024
★★★★★ 4
Good for body, reapply often, use something else on face
Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen
We used this sunscreen at the Oregon coast on a cold but sunny February weekend and it had good sun protection. It’s non greasy and applies easily. No white cast when fully rubbed in and absorbed.
My kids were able to use it without any irritations but I could not use it on my face, it irritated my skin immediately and I had to wash it off with facial cleanser and apply a sunscreen meant only for faces.
I have sensitive skin and some rosacea, so this one I kept to only using on my arms and chest.
One of my kids still got a sunburn on her cheeks after I applied this sunscreen, she had quite pale, sensitive skin, so again I would still recommend using something else on the face.
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Reviewed in the United States on February 20, 2024
★★★★★ 3
Clear but scented
Style: Kids SPF 50 Mineral Sunscreen Lotion, Size: 3 Ounce (1 Pack) Mineral Sunscreen
We go through a lot of sunscreen during summer with little ones, so we’ve tried many brands. This bottle is pretty small, which is both a plus (compact for traveling and putting in your purse) and a minus. I was so pleasantly surprised that this mineral sunscreen doesn’t leave a heavy white cast, as most others do. But, it is fragranced, which is both surprising and disappointing to me for more natural kids’ sunscreen. I do think it is effective, so if you don’t mind having some fragrance, then I would recommend. Our family doesn’t love added fragrances so this isn’t a great fit for us.
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Reviewed in the United States on April 17, 2024