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Description
Rat CASP12 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 a caspase 12 (CASP12) capture antibody. After incubation and washing, the sample is developed 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 12 (CASP12) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Caspase 12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Caspase 12 (CASP12) is a protein encoded by the CASP12 gene. This protein belongs to a family of enzymes known as caspases, which cleave their substrates at the C-terminal aspartate residue. It is closely related to caspase 1 and other members of the caspase family, known as inflammatory caspases, which process and activate inflammatory cytokines such as interleukin 1 and interleukin 18. This gene is polymorphic, producing either the full-length caspase protein (Csp12L) or an inactive truncated form (Csp12S). | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.7 ★★★★★
Based on 499 reviews
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Product Reviews
★★★★★ 5
My dog loves this toy!
Color: Green
I was NOT expecting my dog to like this toy, but she loves it! The legs are stretchy and the body vibrates. The battery lasts about a month.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 8, 2026
★★★★★ 5
Great interactive toy. Worth the price.
Color: Blue
Our Lab absolutely loves it. She throws it all over the house. It’s so fun to watch her play with it. It’s by far her favorite toy right now. It’s easy to charge and came with charging cable. It has a button to turn off and on and a couple different speeds. My parents got her a different one for Christmas and since she loved it so much we decided to get her another one. It’s pretty durable too. She normally rips through toys pretty quickly and both of the ones she has are still in good shape. No rips or tears. She hasn’t tried just chewing on it like she does her squeaky toys. She just throws it all around the house and chases it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 17, 2026
★★★★★ 4
Fun, enticing dog toy. As stated in description it won’t hold up for aggressive chewers
Color: Blue
Rating this based on description. My dog loved this toy. It was smaller than I had pictured in my head, but that was on me. I did read the description ahead of time, which clearly states this is not for aggressive chewers. My dog is trained not to rip toys apart so I wanted to give this a shot, but i guess just her jaw strength and teeth were too much. She carried it everywhere. I turned it on periodically. The movements are small, but enough for her to be captivated with the toy. It did mostly survive for about a day before I needed to take it away due to the Velcro opening. Once it started to pull apart she did chew a piece of the Velcro off. It might be helpful with a flap over design covering the Velcro. I have a singing Santa decoration like this where the Velcro is covered up and I have to pull it back a little to get to the Velcro opening. The gaps on the sides of the closure are a bit to large making it tempt my dog to pull and open it up.
I wish there was a more durable option that could hold up to aggressive chewers, but this product was clear about that in the description. I think the opening to the batteries could use some improvement. Granted I do not have the perfect answer to fix this, but I am also not a toy designer.
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Reviewed in the United States on April 19, 2026
★★★★★ 5
A Toy Ball for All Seasons!
Style: Small Ball, Size: 1 Count, Style: Small Ball, Size: 1 Count
Oh my gosh! I’m so happy to find the almost exact ball my 5 1/2 pound 9 month old Morkie, Charlie, that mommy lost at the campgrounds! It was his favorite toy and although he has others, he loved this one above all others, so to find another one was an answer to my prayers. The difference is so minor and in a good way. It is very durable as Charlie has very sharp teeth, but this one is just a bit softer making it easier to hang on to playing fetch. But for Charlie, the size is the same, the squeekiness is the same, the color is the same (yes they can see some colors) and he went into a jumping frenzy when I took it out of the package and the price was unbelievably inexpensive for such a high quality toy! Thank you! Thank you! Thank you! Good mommy! ( I’m ordering another one as a back up, jic!)
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Great investment.
Style: Small Ball, Size: 2 Count
Dogs love them. Durable. Chew worthy for a beagle and a German shepherd. You cannot smell the flavor imparted but the pups definitely can. Always purchase the amount you need so each dog can have one.
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Reviewed in the United States on May 19, 2026
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