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Description
Mouse C3c 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 a capture antibody against Complement C3c (C3c). 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 C3c (C3c) 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 C3c ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement fragment 3c (C3c) consists of three peptides: the C3c β chain (23-667), C3c α chain fragment 1 (749-954), and C3c α chain fragment 2 (1321-1663) linked by disulfide bonds. The C3c component is central to both complement activation pathways and is cleaved by different, specialized proteolytic systems to form the C3 convertase. C3 cleavage releases C3a and C3b fragments, the latter of which is part of the alternative C3 convertase. C3 levels can be very low due to decreased synthesis or consumption. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 3
Great slippers but disappointing quality.
Size: 7.5-8.5 Women/6-7 Men, Color: Ash, Size: 7.5-8.5 Women/6-7 Men, Color: Ash
Extremely comfortable, attractive slippers but the stitching came loose on both leather trim areas almost immediately. I tried to sew/glue down the stitching (as seen in photo) but it kept unraveling. Disappointing as these slippers are great in every other way (though they do run a bit big...I wear a 9 but ended up with a 7.5-8.5 which fits with a bit of room to spare even with thick socks). I just hope they hold up after my latest attempt to stop the thread from pulling out any more as I do really love wearing them.
Perhaps I just got a bum pair?
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Reviewed in the United States on April 3, 2024
★★★★★ 5
Tight at first, but they stretch after a short time...feel perfect after the first week!
Size: 9.5-10.5, Color: Charcoal Faux Fur
I love them!
My first impression was "oooh, this is too tight". I wear a size 10 shoe and honestly, the 9.5-10.5 felt tight when I first tried it. Without socks it felt okay, with socks my toes were right at the top. But I loved the style so much I decided to give it a chance - I am SO glad I did!!!
They do stretch and give, so don't expect it to be tight forever. I have had them a week or so, wearing each day (I WFH) and they feel really nice. I also live in the mountains, we just had our first winter storm, and these slippers are nice and toasty. Exactly what I needed.
The memory foam is plush, though I have had other slippers with it and I know it will squash down over time. Right now, it is a nice feeling as I walk around the house. They are super cute looking, wearing leggings allows me to feel like I am stylish even while still wearing my sleep thermals. YAY!
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Reviewed in the United States on November 18, 2025
★★★★★ 5
favorite slipper ever
Size: 8-9, Color: Charcoal Faux Fur
I love these slippers. They look great with pants tucked in. Incredibly warm and comfortable. I wear them almost every day. Fit is true to size. They stay up but are not tight at all. On top of that I get compliments on them. Highly recommend.
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Reviewed in the United States on May 3, 2026
★★★★★ 5
Favorite Slippers Of All Time
Size: 6.5-7.5, Color: Charcoal Faux Fur
ETA 7/2024: So I ended up buying a size 5-6. They fit better around the top and sides of my foot but the sole is a bit small. The 6.5-7.5 sole fits better but the material around the top and sides is roomy so it bunches up. Sigh. I still love these slippers but the fit is frustrating.
I'm obsessed with these slipper boots! They are so cute and comfy.
They look especially good with leggings but I also like that I can tuck other types of pajama pants into them. I'm short and have to fold the waist of most pajama pants so they're not super long on me, but with these boot slippers on I don't have to worry about my pant bottoms dragging on the floor since I can just tuck them in.
I like that they have rubber soles. I love the look of the faux fur but not sure how easy they will be to wash because of it.
I wear a 7 or 7.5 and I got the 6.5-7.5 size. Even with thick socks on they are roomy. Because they are house shoes it doesn't matter much but I probably could've sized down.
I like the way they look so much I might buy a size down and use that pair for wearing outside of my house.
Some other reviews said they have a strong smell but mine did not.
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Reviewed in the United States on January 1, 2024
★★★★★ 4
Cozy warm, overall a good slipper!
I ordered a size Medium and I normally wear a women's size 8 US / 39 EUR and they fit well. According to the box, a Medium in Acorn sizing is Women's 6.5 - 7.5, but I do think these run a little large which is why I went with the Medium vs. Large. The boot part fits well, not too snug or too loose, and it stays up if it has something to adhere to (like over pants), otherwise it would slide down my bare leg. I am super sensitive to scratchiness lol, so I keep the boot part over pants because it is a little scratchy to me. These slippers are nice and cozy, so they keep my feet / ankles warm. The footbed provides a nice cushion and the soles/suede guards are strong but without making you feel like you are wearing actual shoes / boots. I have noticed that the footbed does not have a lot of rebound so there are definitely impressions of my toes inside... which means no one else will want to wear them... haha! I've also noticed the seams seem prominent on the inside, I wish they were a less noticeable, but that is just one of my hangups. So, the only reason I am giving a 4 instead of 5 is the inside is not as soft as I'd like and the seam problem. Regardless, I wear these all the time at home (which is where most of us all are right now) and I am happy with them overall. Oh, and I got the Buffalo Check pattern and it's super cute and fun!
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Reviewed in the United States on December 5, 2020
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