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Description
Human CALCOCO2 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 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, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a calcium-binding and coiled-coil domain-containing protein 2 (CALCOCO2) capture antibody. 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 calcium-binding and coiled-coil domain-containing protein 2 (CALCOCO2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader, and the sample concentration is calculated. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Calcium-binding and coiled-coil domain-containing protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Calcium-binding protein coiled-coil region 2 is a protein encoded by the CALCOCO2 gene. The protein encoded by this gene is a subunit of the nuclear domain 10 (ND10) body. ND10 bodies are nuclear domains that appear as ten small dots per nucleus on immunohistochemistry. Based on their resistance to nuclease digestion and salt extraction, they are thought to be associated with the nuclear matrix. ND10 proteins are removed from the nucleus following HSV-1 infection and may play a role in the viral life cycle. Diseases associated with CALCOCO2 include granulomatous disease, chronic, autosomal recessive, and salmonellosis. Gene ontology annotations associated with this gene include protein homodimer activity. An important homologue of this gene is TAX1BP1. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Great security features.
Size: 1 Camera
Works great I just wish the battery last longer.Nightvision is great as well weather proof.Sticks up and is sturdy as well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2026
★★★★★ 5
Great Product
Size: 1 Camera
Great camera for outdoor security. The features on the app have been updated and work better than the Ring Doorbell.
The camera has a great live picture, and the sound is very clear.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026
★★★★★ 5
Quality Cameras
Size: 2 Cameras
Love these Cameras. Great clear quality and super easy to set up! If you are looking to add a camera or camera system, you can't beat the price and quality of these
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 18, 2026
★★★★★ 5
Read this if you're having connection or audio/video/notification difficulties ***UPDATED at end***
Before I start my revised review, let me state that there's been a lot of news lately about people hacking into Ring cameras, but this only happens when someone installs a new camera and doesn't update the default username and password (or has an extremely weak password which is easy for a hacker to guess). This is like leaving your front door open while on vacation. Your camera is sending and receiving signals over a wireless network. Anyone within range of the signal can potentially be a threat if you don't have good encryption (username and good password) set up on your camera. There's no logical reason for a Ring owner to not do this. Ring even stresses changing the default settings during the initial setup process. Bottom line, don't blame the car manufacturer if you leave the keys in your car and it gets stolen.
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Recently, I wrote a rather scathing 1 star review about the problems I was having with my new Ring Stick Up Wireless Battery Cam. I'm no stranger to wireless cams, having 10 hooked up and running seamlessly on another network as I write this, but I was totally frustrated with the connection reliability, not only with my two upper end cellphones, but also on my Galaxy Tab and Amazon Show. More times than not, the software wouldn't connect with the camera but when it did, the video was choppy. Notifications were also hit and miss. I have a great wireless mesh router system throughout the house, so after doing all the troubleshooting I could think of, I assumed it was due to faulty hardware or software. I gave up and started the return process through Amazon. It was a new product after all and new products sometimes are really buggy. Plus, I read other negative reviews which seemed to justify my thoughts and actions.
Then I got an email from a gentleman from Ring saying he read my review and asked if I'd be willing to let him help troubleshoot my issues with me. Although I had already started the return process, I was willing. It was refreshing to know Ring Customer Service was genuinely interested in troubleshooting possible defects in their newest product.
Together, we isolated the issue within about 20 minutes. It turned out it wasn't the camera or software after all.
Anyone having connection issues should do what he advised me to do. Open up the Ring app and click on the 3 little dots to the upper right of the camera screenshot, then select settings. From there go to Device Health and check the signal strength. Although I had a good mesh hub (inside) within 10 feet of my camera (mounted outside), my signal strength was RSSI -64. He stated that the signal was so weak it was on the cusp of what is needed for the camera to even minimally operate. This prompted me to reboot my wireless mesh system and reset/reconnect the camera. When it came back online, my signal strength went to -39, a vast improvement.
A quick check of all of my devices showed the camera and software working flawlessly. Hat's off to Ring.
Anyone who may be having connection or audio/video/notification difficulties, check your signal strength to the camera via the Ring software, even if your router is close by. You just might be surprised.
***UPDATE***
Camera (and software) is still working flawlessly on all devices.
Ring has now updated their Windows 10 desktop app (which previously didn't work with the new wireless battery cams) and it works flawlessly as well.
Observations.
Notifications on my phone, Tab and Amazon devices (Show, Echo, Dot) are almost instantaneous. Alexa says "Someone is at your front door." Very cool.
I bought an extra battery, but it looks like it'll be at least a few months before I need to swap it out, even with the intensive testing I've done on the Stick Up Cam. Battery life seems excellent.
I have one zone blocked out and that seems to work fine as well. I also have it mounted a couple of feet above and to the left of my front door, tilting downward at an angle. Even though residential traffic is visible in the field of view, I've not gotten one trigger due to street activity. The motion sensor is triggered by anything coming into the bottom half of it's field of view. From the time someone triggers the motion sensor until the time I'm aware of it is 1-2 seconds. That's about as close to real time as you can get. It's not given me a false alarm even once.
Telling my Amazon Show to "(Alexa), show front door cam" takes about 5 seconds to complete.
I can also watch triggered video clips and get a live feed while away from home without any extra setup to my network. It's totally automatic.
Because of the customer service I received from Ring and the fact the product is much better than I originally thought, I will be expanding my Ring family soon.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 29, 2019
★★★★★ 5
Reliable camera with solar keeps it running
Size: 1 Camera
I’ve been running this camera with the solar panel and it’s been solid. The video quality is clear day and night, and the motion alerts come through quick without a lot of false alarms. Setup was straightforward and didn’t take long.
The solar panel has been a big plus. It keeps the battery charged so I don’t have to mess with taking it down to recharge. As long as it gets decent sunlight, it stays topped off.
The app is easy to use and lets me check things anytime without issues. It’s held up well in the weather so far. Overall, it does what it’s supposed to do and gives me peace of mind. I’d recommend it if you want something low maintenance.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 14, 2026
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