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Description
Human C3 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 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 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 Complement Component 3 (C3) capture antibody. After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Complement Component 3 (C3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Complement Component 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement protein 3 (C3) is a protein of the immune system. It plays a central role in the activation of the complement system and contributes to innate immunity. Organisms deficient in C3 are susceptible to bacterial infections. Several crystal structures have been determined, revealing that the protein contains 13 domains. The C3 precursor protein is first processed by removing four arginine residues, forming two chains, β and α, linked by a disulfide bond. C3 convertase activates C3 by cleaving the α chain, releasing C3a anaphylatoxin and generating C3b. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.2 ★★★★★
Based on 2272 reviews
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Product Reviews
★★★★★ 4
Best academy I've read this year
Format: Kindle
I need a few things when it comes to a first book of a PNR romance series
1-Good world building (which this totally did)
2-An FMC I can root for (oh hell yes, Pandora is someone I can cheer for)
3-Good drama (can you say GROVEL BOYS!)
4-Enough story to make you feel like you really read something with meat (you saw this book is like 600 pages, yeah?)
5-A hook at the end so I want more! (please, Lyra, gimmie more?!? I need more!!)
Be aware this book is a slow burn, but damn do I feel like there'll be some big payoff when it finally happens. Who doesn't like the buildup?
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Reviewed in the United States on June 4, 2024
★★★★★ 5
Pandora’s Pain, Power, and Passion
Format: Kindle
I absolutely love this new world Lyra Winters has created! The spin on a Demon Academy setting was fresh, unique, and completely addictive.
Pandora is a character who immediately captured my heart. Thought to be powerless and enduring years of brutal abuse from her mother, it’s no surprise that her powers emerge at the exact moment she needs them most. After her mother’s death, Pandora discovers her father is none other than Death himself, a soul eater with a dark legacy. Her journey at the academy is anything but easy, filled with challenges tied to her father’s infamous reputation, her barely controlled abilities, and the cruelty of those around her.
Pandora is easy to root for, you feel every ounce of her pain, resilience, and growth. Along the way she meets Reed, a half-human dream demon who’s kind, steady, and the kind of friend everyone wishes they had. There’s also Hunter, a vengeance demon and counselor connected to her father, who adds another intriguing layer to her story. Then there are the bullies: Dexter, a brooding shadow demon; Bram, a chaos demon with a drinking problem and deep hatred for demon nobility; and Skel, a fear demon wrestling with his own darkness. They might hurt her, but they also can’t seem to stay away when she’s in danger, making for some deliciously complicated dynamics.
This book hits so many of my favorite tropes: friends to lovers, enemies to lovers, and of course, the irresistible “who hurt you?” storyline. I devoured it, and I’m already diving straight into book two!
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Reviewed in the United States on September 4, 2025
★★★★★ 5
So good!
Format: Kindle
Oh my goodness! What did I just read? Lyra Winters you have some serious explaining to do! That cliffhanger killed me! I am so happy to be back in the world of Kalista. This is definitely darker than Fates Hollow but oh so good. This is a fated mates reverse harem which I absolutely love.
Pandora had a very hard and rough upbringing. She lives in pain constantly and it makes it hard on her. She struggles with everything because she was kept so isolated and is new to her magic. Pandora gets sent to the Reform Academy and all of them have reasons why they are there. I love how, after everything she has been through, she is still a nice person. She is growing and becoming stronger too. Love her character.
The guys all act like jerks at first but all have a back story that helps understand why even though want to smack them. I'm here for the groveling that I'm sure will come. I love them all. They each bring something for her. They are all drawn to her though.
This is a slow burn book but there will be more books in the series so sure it will build. Man, that cliffhanger was a doozy and need book 2 now!
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Reviewed in the United States on June 3, 2024
★★★★★ 3
Good-ish
Format: Kindle
🌶️ 0/5
⭐️ 3/5
I will preface this by saying I am not a huge RH fan. However it didn’t feel like a RH quite yet. The relationships and plot are all building in this book. At first I was thinking things were happening stupid fast since she’s had zero interaction with another being and has been tortured her whole life, and I believe they were for one side of the relationship, but by 50% things moved too slow lol. Idk I think the main 1-3 guys I’m actually interested in more than the others didn’t get enough air time so I hope the next book things start moving with them since it ended the way it did.
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Reviewed in the United States on April 12, 2025
★★★★★ 5
Can I have Term 2 now?
Format: Kindle
This is by far my favorite book of Lyra's to date. I gobbled this book up so fast, I was so upset I finished it. I know this is going to be a book I reread, just like Fate Hollow Academy.
I loved Fate Hollow Academy, so I was excited when Lyra announced she was taking us readers back to Kalista. Although the characters from Fate Hollow are mentioned several times throughout the book, this book is about Pandora and her love interests in the Demon Realm.
Before you deep dive into this book, definitely look at the trigger warnings before you start. When Lyra says there's dark themes in this book, she meant it. Also, just know this book does end on a cliffhanger and will probably leave you with as many questions as it did me.
Below may contain some minor spoilers, so keep that in mind if you want to keep reading my review.
Pandora's first 2 decades of her life are full of torture, hatred, and confinement in a cellar. That is until her father finds her and brings her into the world she was supposed to live in. Because of her upbringing she wasn't acclimated to the Demon world and the way their society works so when she is offered the chance to go to a Demon Reform Accademy, she accepts it to learn the ways of society and how she is supposed to feed.
Pandora isn't like the other common demons in this world, no. She, like her father Death, are soul-eaters. Pandora is too scared to release her powers because she doesn't want to kill anyone, so she needs to learn how to feed off of a piece of soul instead of taking the whole thing.
Because of who her father is, she's classed as a Noble, and while she doesn't agree with her fellow Noble students, who look down on the "lower" class, she also is faced with 3 Demons who hate her for being a Noble. Theses a lot of back and forth tension which these 3 demons who have issues of their own. One is obsessed with her, and wants to know all of her secrets. One leaks fear all over the place, but also is scared of the "princess" -wink- -wink-. One is a complete drunk who doesn't like her just because he's got family Noble problems.
Don't worry, there are 2 other demons who are just the sweetest towards her. One who will get vengeance for anything done to her, he's got Golden Retriever energy around her and touch her and die energy for anyone around her. Finally, there is one who is friends with her even when the whole school is scared of her, who brings her into his dreams and will create nightmares for those who bother her.
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Reviewed in the United States on June 4, 2024
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