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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 79 reviews
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Product Reviews
★★★★★ 3
My opinion
Format: Kindle
Let me preface this by saying I really liked the story and the characters however, this book is in desperate need of some sort of editing. It's not misspelled words or formatting, but continuous run on sentences. Redundancies within the sentences. There were a couple of paragraphs that I had to go back and reread 3 or 4 times. Overall, I'd say it was worth the read.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 29, 2025
★★★★★ 5
Beautiful
Format: Kindle
I love omegaverses I had looked at this one multiple times thinking it's just another book about cocky rock star guys that let fame get to their heads and there are parts like that so I wasn't 100% off. I started reading the second book and met the fmc Astraea from the first one,after she had a run in with the aphla from the gym.😁 it gave me the push to stop and read book one first and im so glad I did this book was amazing their was so many characters I fell in love with hoping they will find their happily ever after. the guys were great, the plot was 🎯, and the ending had me 😭😭😭. I was wondering how nate could ever redeem himself, and he did. the last scene with him was sad, but I also felt it was beautiful. thank you to the author for making a beautiful omegaverse book that gave me all the feels. now I'm jumping straight into book two.
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Reviewed in the United States on March 25, 2025
★★★★★ 4
Knot their Omega
Format: Kindle
It took me a day to really get into the book, there is a lot happening in the beginning of the story. I wish there was more of a time frame because I felt like something’s happened quickly but they were farther away than I thought so that was a little confusing to me. I felt like there was so much happening but also not enough.
As for the story itself I enjoyed it, it was different than other OV I’ve read. Bring the tissues, you will need them. They guys were great, I wish we saw more of Kai and Kenjis relationship. As for Nate…… IYKYK.
I enjoyed this book, my first from this author and look forward to reading about Kamaris story.
Book: ⭐️⭐️⭐️⭐️/5
Spice: 🌶️.5/5
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 24, 2024
★★★★★ 2
Has Potential, Poorly Executed
Format: Kindle
I really really wanted to enjoy this book and was looking forward to reading it. However there were just one too many convoluted plot points, sentences just did not flow nicely at times, and I was left with a lot of confusion on my end. I think it has the bones of being a great story but seems like it was rushed and I had to push myself to finish the last of the book and was left with an unsatisfying ending. It’s definitely different from other omegaverse novels I’ve read but not for good reasons. I really think it could be great but needs some serious tweaking and editing before I’d read it again.
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Reviewed in the United States on February 3, 2025
★★★★★ 5
Secrets, Seduction, and Rockstars!
Format: Kindle
Rockstars, secrets, and off-the-charts chemistry? Sign me up!
Cinder Blaze takes us on a rollercoaster of passion and peril with Knot Their Omega. Blair Vesper, a secret Omega masquerading as an Alpha, strikes a risky deal to tour with Blooming Salvation, a band teetering on the edge of chaos. Enter Icarus Morrigan, the enigmatic manager, and his three complicated and irresistibly sexy rockstars: wild Kenji, icy Kaiser, and fiery Nathaniel.
This book delivers steamy Omegaverse drama, sizzling slow-burn romance, and just the right dash of angst. The tension between Blair and the band crackles like electricity onstage, while the societal stakes add depth to the spicy dynamics.
Short, sharp, and oh-so-sinful Knot Their Omega will have you hooked from the first note!
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Reviewed in the United States on December 9, 2024
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