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Description
Mouse C9 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 uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with Complement Component 9 (C9) capture antibody. After incubation and washing, the assay is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Complement Component 9 (C9) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Complement Component 9 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement component C9 (C9) is a MACPF protein involved in the complement system, part of the innate immune system. Once activated, approximately 12-18 C9 molecules aggregate to form pores in the target cell membrane, leading to lysis and cell death. It is a member of the complement membrane attack complex (MAC), which also includes complement components C5b, C6, C7, and C8. MAC formation occurs through three distinct pathways: the classical pathway, the alternative pathway, and the lectin pathway. The pores formed by C9 are important for killing bacterial cells during infection, and target cells are often covered with multiple MACs. The clinical impact of C9 deficiency is infection with the Gram-negative bacterium Neisseria meningitidis. | |||||||||||||||||||||||||||||||||
| 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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4.1 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Arrived in good condition
Format: Paperback
Arrived in good condition
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 27, 2026
★★★★★ 5
A first Poetry book...
Format: Paperback
Update (7/22/20):
I must elaborate more on my intention for this book, for had my previous brief update felt vague. By practicing writing, I meant understanding and placing down the basic conventions of poetry because I am critical when it comes to writing. As I explore through the pages in this book, involving just the underlining, the structure of each topic in this book makes for ease of learning. Near the end of each topic contains ideas for writing, allowing one to utilize the content for practice. After all, I'm not an expert, just one who has the curiosity for crafted writing. Maybe this book may be sufficient enough to hone in the skills of poetic writing; for me, exploring other poetry books invites more perspective and this is a starting point.
(Book condition)
I have just started reading this book, so there is not much input I can include. For now, my review is not based on the content, but rather the condition of the book. I was bummed that this book came in damaged on the lower spine, and had to order another copy, so I can return the other copy later. It is disappointing, books that have been returned have not been inspected. Even if we cannot contact the representatives, I decided to slip a note about it, so others do not receive the copy I returned.
end of rant.
I will update on this book, once I have immersed myself in this book. I am a beginner and even though I learned poetry in high school, I wanted to delve into this book, so I can practice writing.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 15, 2020
★★★★★ 5
Two practing poets share guidance and inspiration
Format: Paperback
Normally I'm suspicious about guides to writing poetry. The creation of a poem is such a personal and private affair, that I tend to despair of long distance help. Happily "The Poet's Companion" strikes such a balance between inspiration and craft that it's the next best thing to participating actively at an outstanding workshop. I benefitted from the poetic themes and supported exercises, feeling creatively rejuvenated by the authors' challenges. It seems to have no ideological axe's to grind-its only goal is to advance the poetic impulse from the brain to the page. I am truly grateful to have this book as it has aided in a personal liberation from a massive case of writer's block. I recommend this book to everyone of us who loves to read and learn the practices of writing poetry.
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Reviewed in the United States on May 10, 2014
★★★★★ 5
Yay!
Format: Paperback, Format: Paperback
Got the book, plain simple. In tact, no blemishes or damage, no broken spine, nothing. Thank you!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 5, 2024
★★★★★ 5
Like
Format: Paperback
Item was of optimal quality. Notably fascinating packaging. Select delivery. A++!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 20, 2021
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