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Description
Rat CST ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 utilizes 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 corticosteroid (CST) capture antibodies. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the corticosteroid (CST) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Corticosteroid ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Procortistatin is a protein encoded by the CORT gene. Its 105 amino acid residues are cleaved into cortistatin-17 and cortistatin-29. Cortistatin-17 is the only active peptide derived from the procortistatin. Cortistatin is a neuropeptide expressed in inhibitory neurons in the cerebral cortex and shares strong structural similarities with somatostatin. Unlike somatostatin, it enhances slow-wave sleep when injected into the brain. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.5 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
K and N is the best
Size: CA12290-Premium
In my opinion, you can’t buy a better air filter. Been buying them for all my cars since the 1990s.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 8, 2024
★★★★★ 2
Does not fit 2023 Honda Accord as it says in the title
Size: CA12290-Premium, Size: CA12290-Premium
Does not fit 2023 Honda Accord as it says in the title. Seems like good quality otherwise.
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Reviewed in the United States on October 3, 2024
★★★★★ 5
Fits Toyota 2010 FJ cruiser
Perfect fit for a Toyota 2010 FJ cruiser. Install might have been the easiest cabin filter I've done in any car that new. Remove glove box, pull out the cover and replace it. Fits great and takes odors out from it sitting for 6 months in the driveway. No notice to less airflow when on any level of fan speed.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
Comparison with POTAUTO filter: Very similar but cheaper
I bought a POTAUTO MAP 1033C and EPAuto CP846 cabin air filter to compare them for use in my 09 Legacy (gen 4). They both seemed comparable and are cheaper than most other, similar filters, though the POTAUTO was and still is ~33% more expensive than the EPAuto.
Both seem built well-enough, considering they're only being used as relatively low-flow cabin filters. That said, the EPAuto is slightly better, mainly due to the white trim piece being unattached along one side on the bottom of the POTAUTO filter. Almost certainly nothing that will affect its performance or longevity, but it is interesting considering it's the more expensive of the two. However, it must be kept in mind that this is an incredibly small sample size.
In one of my very scientific tests (/s), I held them up side by side and looked through them toward the sun (obviously being careful) to judge thickness/density and uniformity. Neither had any thin spots that I noticed, and they were pretty similar overall. One of them blocked slightly more light than the other, indicating more filtration, but I unfortunately don't remember which one. I feel like it was the EPAuto, but I don't really want to speculate as I could very well be wrong. What I do remember is that the difference was so minor that all else being equal, it wouldn't justify the cost difference between the two. In other words, even if the POTAUTO were the slightly better one, it wouldn't be worth the extra few dollars for the minimal amount of extra filtration.
In another test, I compared the filters to each other and the old filter (which I'm pretty sure was OEM, but certainly not a charcoal filter, so it was significantly thinner) by blowing air from a compressor through them. I held the nozzle at roughly the same distance from each on one side of the filters, and I held my other hand at roughly the same distance from each on the other side. The old filter, unsurprisingly, let much more air flow through. Both charcoal filters were much more restrictive due to their extra thickness, leading me to feel much less air coming through. Both were roughly the same. Both filters also held up just fine to the strong blasts of air.
I bought a couple other filters that I was going to cut to fit to use one or both with these filters as a pre-filter and/or additional charcoal layer. After the airflow test, I decided against this, as these are a lot more restrictive than OEM already, and I didn't want to push it, since that could at best cause issues with getting good airflow into the car, and at worst could damage the blower. If not for the fact many, many people have been using these and similar filters for a long time without apparent issue caused by this, I would hesitate to even use these. I haven't noticed a decrease in the airflow, but it's doubtful I would since I rarely turn the fan up past the first couple settings (usually have it on the first) if I have it running at all, and I have the center vents pulled out (to access the inside of the dash) which causes the flow at the vents to be reduced slightly.
TL;DR - Both the POTAUTO and EPAuto charcoal filters appear to be a good choice, with the EPAuto having a slight edge on build quality (based on my limited sample size of one each) and a cheaper price. Filtration appears to be very similar between the two, certainly not enough of a difference to warrant the extra price for the POTAUTO over the EPAuto. Flow is significantly more restrictive than OEM filter but doesn't appear to be an issue. I give the EPAuto 5 stars and the POTAUTO 4 stars, only because the value of the POTAUTO is a good bit less (very similar or possibly even inferior quality for 33% more money). I can't speak to their longevity or performance, but I don't imagine either should prove to be an issue.
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As a side note relating specifically to the Legacy: replacing the cabin filter in this car is a PITA. It's not overly difficult per se, but a serious pain and certainly not something you're going to do when you have a spare few minutes. I'd rate it probably around a 3.5/10 in difficulty and a 7/10 for annoyance. While you can sort of access it by removing the manual compartment, you can't remove the tray through that. So you need to actually take the whole glove box out, which requires removing the side panel, unhooking the string/loop that keeps it from falling all the way down, and removing a few plastic screws, which can be a bit of a pain (and apparently Subaru loves them since they're all over the car). A stubby Philips driver will be helpful. Once you have the glove box out of the way, you have to unscrew several more of those plastic screws to remove the plastic cover between the glove box and the filter. This hole is where you gain access. Be careful when removing the old filter as loose dirt and debris may fall out and make a bit of a mess. You don't really want to get any in the fan below it if you can help it. Reverse the steps to reassemble it, and remember to reattach the string. Getting the glove box back in its track can be a bit of a challenge; in my experience from doing it multiple times I've found you sort of half force it and half don't. That is, it'll likely offer some resistance even if it's lined up, so if you try to baby it you'll probably be there a while, but also play with the alignment a bit to see if you can get it without marring up the tab and the slot on the right side too much. All in all, expect to spend anywhere from 15-45 minutes on this, and make sure you have a standard length as well as a shorter or stubby Philips screwdriver. I have to say, when it comes to air filters, this car is horrible. The air intake filter is a pain to change, too--much worse than most if not all other cars I've done.
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Keywords: Subaru Legacy, fourth gen, fourth generation, 4th gen, 4th generation, 03, 04, 05, 06, 07, 08, 09, 2003, 2004, 2005, 2006, 2007, 2008, 2009
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Reviewed in the United States on September 29, 2017
★★★★★ 5
Good price on the part and 5 min install saved me $35
Fit fine in my 4th gen 4Runner took 5mins to install
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Reviewed in the United States on April 19, 2026