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Description
Human CRT 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: 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: 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. 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. 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. 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. 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. 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 calreticulin (CRT) 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 calreticulin (CRT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Calreticulin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Calreticulin (CRT), also known as calregulin, CRP55, CaBP3, calsequestrin-like protein, and endoplasmic reticulum-resident protein 60 (ERp60), is a protein encoded by the CALR gene. Calreticulin is a multifunctional soluble protein that binds to Ca2+ ions, inactivating them. It binds with low affinity but has a high capacity, allowing for release in response to signals. Calreticulin is located in storage compartments associated with the endoplasmic reticulum and is considered an ER-resident protein. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
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Product Reviews
★★★★★ 5
I wish I had Siless 20 years ago!!!
Model: SLH20025, Model: SLH20025
I have installed all kinds of insulation in vehicles over many years. I only needed to purchase 2 boxes to complete the entire interior siding of my 2025 chevy express, what a great value! I wanted to insulate my locksmith van for sound and heat. The sound reduction virtually eliminated the road noise. These panels also look amazing. It got up to 110F in the sun with the doors closed. After adding Siless insulation the temperature maxed out at 97F. That is a massive difference. Plus, I didn't even finish with the roof before testing. Siless Hybrid 3in1 was extremely easy to cut and adhere onto metal panels. I'm impressed with the quality of the individual panels and the adhesive is not toxic smelling compared to some others. My favorite thing about these panels is you can actually take them off and readjust them before they finally stick and they do not leave any black residue behind when you pull them up to adjust them. All I needed was a heat gun, knife and a roller to complete this job. The size of each panel is perfect, you can handle them with ease. I probably won't need your product again for a while, but when I do I will definitely be back.
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Reviewed in the United States on September 21, 2025
★★★★★ 5
Works well and is easy to install.
Model: SLH20025, Model: SLH20025
I was sound and heat insulating a generator bay in my class A RV so I did three tests before and during material application: I did a sound level test to see how effective the material was for my application, a "stickiness" test, and a flammability test. This was indoors on a dry 78°F day. From beginning to end, the entire process, including testing, took 5 hours to install 25 sqft or material in a very complex area. One hour for cleaning, 3 hours for installation, and 1 hour for test setup and testing.
Using a sound meter, I took before and after readings and determined that I reduced the sound level by 2.2 db (40% energy decrease) with a noticeable decrease in sound and frequency. The generator bay is located under the master bedroom in the aft portion of the coach. The sound went from a rattling, vibrating generator noise to a more tolerable low level rumble.
The material was easy to apply. I did some testing prior to application and determined that surface preparation is key. I cleaned my painted metal surface and then degreased with acetone and the material stuck well. Be careful and purposeful when you apply the material because once it was stuck to the metal, it was quite difficult to remove or reposition.
Since it was going to be installed into a gasoline powered generator bay located under our sleeping quarters, I did some flame testing to determine the level of risk. Although not highly flammable after installation, my testing revealed that the PE used will catch on fire under certain circumstances. In order to reduce the risk, I used aluminized duct tape to ensure that all the edges were sealed and the PE was not exposed to direct flame in the event of a fire. I also decided to install an automatic fire extinguisher as an added precaution. In all fairness there should be one installed in the generator bay anyway.
All in all, I am very satisfied with the product and the results. I have recommended this product to my other RV owner friends.
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Reviewed in the United States on April 13, 2025
★★★★★ 5
Great product. Best value i could find
Model: SLH20025
I'm using talk to text, so forgive any grammatical and spelling errors.
I ordered this product after doing a bit of research on sound dampening materials. I had been doing car stereo for quite some time since my teens. Itook a break for 20 years or so. I'm now 40 and back in the day there was dynamat. And that was about it. as irecently bought a new 2017 F350 crew cab. I wanted to put in a new stereo system and being that it was my dream truck. I was going to put in my dream stereo. But before I could do that, I needed to start with the sound deadening. I looked into today's leading product and found a company called resonx. Had taken their place as king of the hill in sound deadly. I researched their products and found the materials they were built from and Silas was the closest match at the best price I used Silas Max. First, then Silas 3 and 1 hybrid over that on all exterior panels. Then, on the interior panels. I used the 3 and 1 hybrid . on the floor I used Silas Max. Then, the 3 and 1 hybrid, then Silas liner over that the car is incredibly quiet . As far as sound dampening and deadening, it works very well for the money really can't beat it. Do your homework no How and where to apply it? And you really can't go wrong or about $400. I did. What would have cost me easily? Two thousand in other brands highly recommend the product prep work.Is everything do your homework?Do it right and you will be happy with this stuff
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Reviewed in the United States on May 6, 2026
★★★★★ 4
Not simple, nor easy
Model: SLH20025
I wanted to add sound proof material to the bare metal areas on the floor of my Toyota Tundra. The area behind the front seats had a haphazard application of sound deadening material. There were several complications to the project which may come up in other vehicles.
First is the floor covering which had plastic pillars underneath to support any weight on the cover. The sound deadening material had to be cut out so these pillars would rest on the bare metal of the floor. This required a lot of cutting with a utility knife and at times heavy scissors. The instructions say to make a template before cutting the material and I did this for the driver's side. This doubled the amount of time needed to install it. On the passenger's side I laid the rectangles of material with the backing on and pressed the floor covering onto the material. This showed where the support pillars were, which I then cut out.
Second is peeling the backing off the material. This can be very difficult as the adhesive sticks to it. The backing came off in multiple pieces and sometimes took 5 minutes for one piece. They say to wear gloves when handling the material but this is impossible when trying to get the backing off.
Third is pressing the material onto the metal. The instructions say to use a roller, but this was impossible for the irregular surface I was working on. The hand is the best tool to press the material on.
The material seems to be of high quality and I expect it will reduce the road noise in the cab.
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Reviewed in the United States on May 18, 2025
★★★★★ 5
Bringing a 1976 MG Midget to the modern world!
Material Type: 157mil Closed Cell Foam, Style: 157 mil 36 SqFt
I've been restoring a 1976 MG Midget barn find. I bought some Sinless liner to help make the car rattle less and feel more solid.
This product is easy to cut, easy to install, and extremely sticky. I used it to line the trunk, the interior, and even some under the hood of the old MG. The adhesive stands up to the brutal temperature of the engine bay, and when (by accident) some fuel spilled onto the foam, it didn't fall apart or turn to mush.
Thanks to Siless, the MG now feels like a modern car. The floor pans feel much more solid, the doors make a heftier thud when you close them, and the trunk no longer sounds like a percussion instrument when you touch it.
If you know what you're doing, this product will transform the feel of a car. I found that 36 sq ft was enough for my MG Midget, but for a bigger car I would want more. The pattern on the back helped me to cut perfectly measured squares and rectangles. The foam itself doesn't have a strong odor, it works super well, and has held up to several months of changing temperatures (in the Midwest US.)
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Reviewed in the United States on November 11, 2025