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Description
Human C4BPb 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 2000 pg/mL). Then dilute to the following concentrations: 2000 pg/mL, 1000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.25 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 2000 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000 pg/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 utilizes 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 C4 Binding Protein Beta (C4BPb). 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 C4 Binding Protein Beta (C4BPb) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human C4 Binding Protein Beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | C4 binding protein β (C4BP) is a protein complex involved in the complement system, where it plays an inhibitory role. C4BP has an octopus-like structure with a central stalk and seven branched α-chains. The predominant form of C4BP in human blood is composed of seven identical α-chains and a unique β-chain, which in turn binds to the anticoagulant, vitamin K-dependent protein S. C4BP is a large glycoprotein (500 kDa) with an estimated plasma concentration of 200 μg/mL and is primarily synthesized in the liver. The genes encoding the C4BP α-chain (C4BPPA) and β-chain (C4BPB) are located in the regulator of complement activation (RCA) gene group on the long arm of chromosome 1, adjacent to other complement inhibitors. | |||||||||||||||||||||||||||||||||
| 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 | 31.2-2,000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.8 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Fun tug toy that holds up well
Size: Dog Rope Toy - M
We got this rope tug toy for our dog and it’s been a hit. He loves playing tug of war and will happily drag this around the house looking for someone to play with. The rope is thick and feels like real cotton, not that super cheap stiff stuff. It has some give to it but still feels sturdy.
So far it has held up well to lots of chewing and tugging. Our dog isn’t a hardcore destroyer, but he definitely tests his toys, and this one is still in one piece. It’s also a nice size, not tiny, so it works well for medium and bigger dogs, but a smaller dog could still enjoy it.
Overall, it’s a simple toy that does what you want it to do. Keeps the dog busy, gives them something safe to chew on, and holds up better than most rope toys we’ve tried.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 3, 2026
★★★★★ 3
Red flag!!!
Size: Dog Rope Toy - M
Within an hour of giving this toy to my dog, she chewed off the plastic handle and the plastic ball. The worst part is that they came off in very small pieces, some like crumbles. This could have been a serious problem if I had not been watching her and able to take it away.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 12, 2026
★★★★★ 4
Hoping for the Best
Size: Dog Rope Toy - M
Seem pretty strong, but I haven't had that long - hope it holds up to my chewer
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 14, 2026
★★★★★ 5
great dog toy, would buy again
Size: Dog Rope Toy - M
Good value and good material. Great size and thickness. My pit loved it until I drove over it with my riding lawn mower. I will be buying another soon.
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Reviewed in the United States on May 15, 2026
★★★★★ 2
Updated title: Not for a power chewer
Size: Dog Rope Toy - M
Update: I had to cut the ball off the same day I wrote this review. Two days later, the yellow rope was totally unraveled and I had to throw it away. She got maybe 3 days of play with this toy. Do not buy this toy if you have a larger dog that chews a lot.
I will say our dog loved this toy the moment she started playing with it. But the toy doesn't last long with our superchewer. I was hoping to use it for some games of tug, and while we're at it get her teeth cleaned by the spikey red ball.
The listing includes "large" dogs. My large dog (11 m/o Bernese puppy) had this toy broken within the first 10 minutes. The red pieces are a pretty lightweight plastic that she just broke apart completely, specifically the bone on the "handle". The ball is lasting a little longer, but she put a hole in it. We will probably need to cut the ball off as well because she'll chew it up and swallow it if we're not careful, and then she'll need (another) surgery.
The positive is that she loves chewing on rope and the rope pieces seem pretty sturdy. We still play tug with the ropes, but this one I will probably just give to her as a chew toy. We found a more durable ball on a rope toy at Target for a little bit less.
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Reviewed in the United States on July 14, 2025
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