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Description
Human CASP8 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, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a Caspase 8 (CASP8) capture antibody. After incubation and washing, color development is performed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Caspase 8 (CASP8) in the sample. Absorbance (OD) is measured at 450 nm using a microplate reader to calculate sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Caspase 8 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Caspase 8 (Caspase 8) is a caspase protein encoded by the CASP8 gene. It most likely interacts with caspase-3. CASP8 homologs have been found in many mammals for which complete genomic data are available. The CASP8 gene encodes a member of the cysteine-aspartate protease (caspase) family. This protein participates in programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with the Fas protein FADD. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
Really supportive, comfortable pillows, great value.
Size: Queen Size(2 Pack)
When I received these pillows, I followed the directions to fluff them. They were delivered vacuum packed. The first night I slept on them, I didn’t remove any of the filling. That was a problem because the pillows are so over stuffed that it was difficult to sleep on them. I was disappointed until I removed about 1/3 of the filling. Doing so made all of the difference. These pillows are very nice. They keep their shape during the night and provide excellent support for my neck and head. They don’t flatten out or separate like regular shredded foam pillows.
They are well made with an outer cooling cover as well as a inner zippered cover for the foam.
I love the fact that these pillows are adjustable. I saved the foam I removed in the zippered bag the pillows arrived in, in case I needed to restore the foam in the future. They are a very good value for the money.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 25, 2025
★★★★★ 4
Not cooling throughout the night
Size: Standard Size(1 Pack)
The quality of the pillow was great the stitching as well. The one side did feel cool, but it did not last throughout the evening. Also, it was too thick for my neck and caused me headaches. The phone pieces that are inside made me concerned that after time they would flatten and the pillow would get firm. I am a hot sleeper and I expected it to stay cool longer than two hours. Because of that I returned it, but if you like thick pillows, and you’re not a hot sleeper, it may work for you.
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Fantastic I highly recommend him
Size: Queen Size(2 Pack)
We have had these pillows for over a year. I can honestly tell you I absolutely love them. They are cool and extremely comfortable. I like them so much I bought a second set for our camper.
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Reviewed in the United States on June 4, 2026
★★★★★ 3
Decent pillow
Size: Standard Size(1 Pack)
Nice pillow. When first opening it you fluff it and put it in the dryer for about 10 minutes, comes out exactly how I like my pillows, firm but has a little bit of softness. Used this pillow for several nights now. Holds it's shape at the start of the night, but when I wake up its sorta flat and had a smashed feel to it. Fluffs back to it's original size and shape. Just wish it was more dense with about 1/4 to 1/2 more foam inside. Would probably help firm it up and hold it's shape better.
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Reviewed in the United States on June 1, 2026
★★★★★ 5
Firm and dense to the Extreme!!
Size: Queen Size(2 Pack)
I love my pillows, extremely firm and also extremely dense. I hate when a pillow gets worn down in the middle after like a few weeks or a month and then you’re struggling to get it full again. It’s absolutely impossible.
When I got these pillows, it said that it might take 24 hours for them to fully enlarge. I enrolled them and took a look at this flat thin thing and I thought, oh my God, I made a huge mistake in ordering these. Went into the kitchen, made some dinner after dinner went into my bedroom and holy moly, these pillows were massive. I was so excited and I couldn’t wait to sleep on them.
Now mind you, I am a side sleeper, so that’s why I like a very full and firm pillow. I do love how full they are, however, it was a little much. Luckily, you can unzip them and take some of the filling out.
A little tip is to keep the little zipped bag they give you and you can put some extra filling in there and zip it and if you ever want to fill it back up, it’s easy. I took about 15% of the filling out of each pillow which to me was perfect, but I could see people wanting to take much more out. I love that you have the option of taking it out and putting it back in if you ever need to, but I don’t think you’ll need to because these pillows will never get squishy or flat. I love these pillows so much that I don’t know if I could ever exist without them. Please never discontinue these because if I ever need to get another set, I want them available.
Best pillows I’ve ever had in my entire life. Period!
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Reviewed in the United States on December 15, 2025
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