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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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4.0 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Wonderful Insight to the Seer Prophetic Ministry…Those Who Are Called To This Ministry Will Be Greatly Benefitted!
Format: Kindle
I just finished reading this book and I thought it was fantastic. It is the first book I've read on the seer prophetic ministry. I was an atheist for years, but then got supernaturally saved and baptized in the Holy Spirit at the same time and immediately began moving in dreams and visions. I thought it was odd that I would mostly receive revelation for myself, others, and the church through dreams, rather than ministering to a person and receiving a word as I was speaking to them. I would pray and ask God to give me revelation through visions as I would minister in person on the spot but it would very rarely happen. I learned through this book that I'm actually a seer and received my calling last year. I then found this book and felt it clearly outlined my gift and helped me to understand who I am and how God made me to function. The seer ministry has never been taught in any church I have attended and I've been to plenty charismatic churches. I find this sad but I believe it won't always be so. There's also an impartation from this book. Before anyone dings me on that, remember that even in the bible, the anointing was transferred through handkerchiefs. I felt this book was pretty solid biblically and James speaks with a heart of love. You will learn more about this ministry and if you're already functioning in it, you may learn some great nuggets and may even be sharpened like I was. I'm looking forward to reading more on this awesome ministry. Thank you James for sharing your wisdom and insight. I'm actually reading it a second time, that's how great it is. Give it a try, you won't be disappointed!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 23, 2015
★★★★★ 5
A GOD-centric UPDATE that guides you to the Throne Room.
I was given the ORIGINAL version of this book as a gift—long before I knew about the prophetic as a realm ALL Believers can enter. I had several false starts reading the book. With each round of attempting to read it I stopped short for the same reason—I felt there was more emphasis on experiences and the author’s revelations than on the Word of God.
Fast forward to several years later, I decided to read this book again BUT by this time I had my own personal revelations and substantial teaching about the prophetic. I was/am still a WORD girl so as I prepared to read it I discovered THIS EXPANDED EDITION and boy was it a wayyyyyyy different read. EVERY IOTA of teaching is anchored in the word of God and turns it all back to the Godhead. There are endless stories of his or other giants’ personal experiences but EVERY account is in the framework of the Word of God. The Godhead is exalted in this book. EVERYTHING else is secondary. I absolutely loved this new writing voice—such that I have since bought another book by the author and I plan to read more of his works.
At the end of the day, heavenly encounters are purposed by God to draw us and others into deeper relationship with Him. The encounters and revelations are not the endpoint, HE is—His Word, His will, and His way. This is the thesis statement of this book and I love it!!!!!!!!!!!!
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Reviewed in the United States on September 15, 2019
★★★★★ 5
Seeing in the Spirit is biblically valid - book helped me understand & lose my fear/confusion about the supernatural
Wow! I read this book over the course of just 2 days. I bought it a while back but hadn't read it until now. I had some fear with seeing in the spirit realm. After reading this book, I have a MUCH greater understanding. I am also now interested in contemplative prayer - which James Goll briefly touches on in this book. I am willing to finally leave prejudices behind about the wonderful and supernatural means that God DOES in fact use to communicate with us. I have bought into the lies that this is somehow of the occult or non-biblical. Further inspection of the scriptures for myself (away from any teachers - no matter how great I thought they were) showed me this is not the case. God DOES speak to us, He DOES manifest Himself to us when we obey and love Him, and there IS a tangible, actual presence of God. What a shame this is being demonized in many Christian circles because there is NOTHING more wonderful and joyous than God's actual presence. Seeing in the Spirit is also another way God interacts with us. I have abandoned my fears and have come to realize how truly amazing this really is.
The Bible is filled with visions, dreams, divine interactions, etc. Any person reading the Bible - setting aside other opinions influencing him/her - will plainly see that the gifts will not cease until the return of Jesus Christ. Also, that we are to hear God's voice and commune with Him. John 5:39 - hit me hard during a terrible time of fear and confusion because of all the various doctrines on this topic. "Search the scriptures: for in them ye think ye have eternal life: and they are that which testify of Me. And ye will not come to Me, that ye might have life." We don't know Jesus through the Scripture, we know about Him from the Scripture. We do have to go to Him and seek Him in addition to studying the scriptures. He is a real Person, and this is a real relationship.
I tabbed the section James Goll wrote on testing the source and testing the spirits as this is something that I have not always done and now have excellent information about from this book. I also have reassurance that this does not offend God and/or Holy angels - as this is what we are told to do. I highly recommend this book - and I think it would be tremendously awesome for more Christians to leave the fear of deception, fear of the devil, and fear of the religious folks ridicule behind. We are able to enjoy a *REAL* relationship with Christ even during our time on earth.
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Reviewed in the United States on July 2, 2016
★★★★★ 5
Good book
Such an amazing book as someone who dreams and sees , it was a great break down and introduction to the world of seeing in which I’ve been looking for . Finished reading it once, now I will read it again
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Reviewed in the United States on August 29, 2025
★★★★★ 5
great book
loved the book
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 30, 2025
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