C4b-binding protein (C4BP) is a protein involved in the complement system where it acts as inhibitor. C4BP has an octopus-like structure with a central stalk and seven branching alpha-chains. The main form of C4BP in human blood is composed of 7 identical alpha-chains and one unique beta-chain, which in turn binds anticoagulant, vitamin K-dependent protein S.C4BP is a large glycoprotein (500 kDa) with an estimated plasma concentration of 200 micrograms/mL synthesized mainly in the liver.The genes coding for C4BP α-chain (C4BPA) and β-chain (C4BPB) are located in the regulators of complement activation (RCA) gene cluster on the long arm of chromosome 1 in the vicinity of other complement inhibitors.
Principle of the Assay
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- C4BPα antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- C4BPα antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the C4BPα amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of C4BPα can be calculated.
Range: 50 pg/ml- 4000 pg/ml
Storage and Expiration: Store at 2-8℃ for 6 months.
Application: For quantitative detection of C4BPα in human serum, plasma, tissue homogenate or cell culture supernatant.
1. One 96-well plate pre-coated with anti- human C4BPα antibody
2. Standard: 0.5ml (4500 pg/ml)
3. Standard diluent buffer: 1.5 ml
4. Wash buffer (30×): 20 ml. Dilution: 1:30
5. Sample diluent buffer: 6 ml
6. HRP conjugated anti- human C4BPα antibody (RTU): 6ml
7. Stop solution: 6 ml
8. TMB substrate A: 6ml
9. TMB substrate B: 6ml
10. Plate sealer: 2
11. Hermetic bag: 1
Please check datasheet for protocol.