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Description
Rat C5d 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 the capture antibody for Complement Component 5d (C5d). After incubation and washing, the sample 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 5d (C5d) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Complement Component 5d ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement fragment 5 is a protein encoded by the C5 gene. It is the fifth component of complement and plays an important role in inflammation and cell killing. Diseases associated with C5 include complement component 5 deficiency and poor response to eculizumab. Pathways associated with this gene include the complement pathway and the immune response/lectin-induced complement pathway. Gene ontology annotations associated with this gene include signaling receptor binding and endopeptidase inhibitor activity. An important homolog of this gene is C3. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
Based on 2167 reviews
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Product Reviews
★★★★★ 5
Love it!
Color: White, Size: 2Tier48in, Color: White, Size: 2Tier48in
Love how this looks like genuine pipes but a little lighter in weight. Sturdy and easier than DIY project!
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Reviewed in the United States on February 26, 2026
★★★★★ 5
Great Commentary
Format: Hardcover
I really like the commentary and expect it to be a cherished addition for my Bible study. The books are well made and should last a long time.
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Reviewed in the United States on November 10, 2025
★★★★★ 5
Font of knowledge
Format: Hardcover
When the package arrived, the box was torn from one side, and I was full of apprehension thinking the books would be ripped, and ruined. Thankfully the books arrived pristine, and no dents or even blemishes.
As for the subject of the books, they are giving me more information than I ever thought I would be getting. These books are excellent to better get an understanding of scripture.
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Reviewed in the United States on August 3, 2025
★★★★★ 5
Excellent Expository Commentary
Format: Hardcover, Format: Hardcover
I recently added the English Standard Version Expository Commentary to my library. This relatively new release has quickly become an invaluable resource for my personal study. The layout is exceptionally well designed. Each section begins with an overview of the passage, followed by the Scripture text itself. A concise section summary and an outline of the verses under consideration then provide helpful structure. Finally, the commentary offers thoughtful exposition, typically presented verse by verse or by clearly defined segments.
Each volume opens with a thorough introduction to the book, beginning with an overview of the text and title and authorship information. Where relevant, the evidence supporting the work’s authorship and reliability is presented, and complex or disputed passages are addressed. The introduction also discusses the date of composition and the historical circumstances surrounding the text.
This is followed by an examination of the book’s genre and literary structure—for example, whether it was written in Hebrew or Greek and why that matters. The author's theological perspective is then explored, along with how the book fits within and contributes to the broader message of Scripture. A key question guiding the introduction is how this book relates to the rest of the Bible. The result is a rich understanding of the cultural context, way of life, and interpretive challenges faced by the reader. An extensive outline precedes the scriptural text itself.
The commentary that follows is highly readable. My older NIV Expositor’s Bible Commentary tends to be more academic, particularly in its treatment of the original languages. By comparison, the ESV edition is both comprehensive and accessible. I consistently come away with all the essential information I need for serious study without being overwhelmed.
Physically, the volumes are attractive and well-constructed. The paper is thicker than that found in many comparable commentaries, the binding is sturdy and appears to be Smyth-sewn, and the typeface is easy on the eyes. The footnotes and endnotes are also clear and readable, contributing to an overall impression of quality and durability. This belongs in any serious Bible student's or pastor's library.
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Reviewed in the United States on January 2, 2026
★★★★★ 5
Very conservative approach, BUT that's not necessarily a hard pass if you're more progressive
Format: Hardcover, Format: Hardcover
A few things you should know if you're considering this purchase:
1. This commentary set has a conservative take on many topics. I haven't read through everything, but that is the general approach.
2. This commentary set is has a very strong complementarian view, and it takes the time to argue this extensively in all the key passages like the Creation story, 1 Corinthians 11, Ephesians 5, 1 Timothy 2 and 3, and Titus 2. It also makes reference to it in smaller passages like Romans 16:1-3 and Acts 18:26.
3. The conclusion given for the practical application of 1 Corinthians 11:2-16 is that this passage should be put into practice with whatever is relevant in the culture you live in. Many denominations would take this approach; very conservative denominations will find issue with the application not being specifically head coverings.
The only reason I'm mentioning these things is so you get a better understanding of what approach this commentary set takes. The fact that I gave this set 5 stars is not because of its conservative views, but rather for the following reasons:
1. The commentary is written in a way that's easy to understand and follow. I felt like the language and sentence structure is pretty down to earth compared to other deeply theological and intellectual commentaries I've read.
2. It's formatted in a way that feels digestible. It goes through the whole Bible taking one chunk of Scripture at a time. So this is what it looks like: A big title saying what the Scripture passage is, then a paragraph or two of Scripture, and then the commentary on the Scripture. The commentary is broken up into these sections: Section Overview, Section Outline, Comment (which has commentary clearly marked verse by verse), and Response. I've included a picture to show what I mean. (Many Scripture passages are longer than the one shown in the picture though.) I find this format to be pretty easy to follow along.
3. It's easy to tell that a lot of thought and effort went into this set. It feels thorough, but not overwhelmingly detailed.
Now, for a bit of my personal opinions about the views this commentary set holds. I'm not certain where I fall in the whole conservative/progressive spectrum. I'm not sure I can clearly define whether I'm complementarian or egalitarian. But I do believe in actively listening to opposing viewpoints, even when it feels uncomfortable to do so. It was truthfully uncomfortable for me to read a very strong complementarian view. However, even in the discomfort, I still came across other things in this commentary set that gave a more clear understanding of Scripture, things I'd never thought of before and that were good for me to read.
If you're more progressive, it goes without saying that this commentary set will not align with your views on everything. However, I don't necessarily think that means the set is absolutely not for you. I believe that the church at large, regardless of where we are on the conservative or progressive spectrum, would benefit from reading and listening to views we might not necessarily agree with. I believe it helps us grow in compassion and understanding and grace toward each other. And even though you may find yourself strongly disagreeing with some of the conservative views, I really do think there's a lot of other commentary that you would probably find really helpful and great to read.
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Reviewed in the United States on August 21, 2025