Encoded by the gene C3 located on chromosome 19, complement C3 is the protein that plays important role in the complement system and contributes to innate immunity. Studies have shown that complement C3 can be purified from human plasma. Here, we provide detailed C3 purification from human plasma protocol for our clients all over the world. In this protocol, a series of steps are designed to remove high-molecular-weight compounds, remove major contaminants, remove hydrolyzed C3, as well as to remove C3 aggregates. Please note that our protocols are only for your reference.
C3 Purification from Human Plasma
Fig.1 Flow chart of C3 purification from human plasma. (Creative Biolabs)
Published Data
Fig.2 Representative chromatograms of various C3 samples resolved by Mono S ion exchange chromatography.1
Native C3 was isolated from human plasma employing in-house purification. Following final processing, the protein was precipitated in low salt, pH-controlled buffer and dialyzed against a 40 mM phosphate solution with ample buffer volume; after 24 hours the solution was refreshed and dialysis resumed for another 24 hours until a visible precipitate emerged. The stock was suspended without centrifugation, aliquoted, and stored at −80 °C, then reconstituted with VB++ upon use. Given complement proteins’ intrinsic lability due to thioester hydrolysis, C3 purity was confirmed by cation exchange and classical hemolytic assays.
As a famous expert with a high reputation in the complement field, Creative Biolabs is committed to the study of various molecular mechanisms and developing a variety of research platforms and protocols. What's more, we are able to provide a range of complementary products and complement assay services, including therapeutic antibodies, serine protease inhibitors, soluble complement regulators, complement component inhibitors, etc.
To learn more details, you can get access with the following links:
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Complement Products
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Complement Test Services
If you are interested in our products and services, please do not hesitate to contact us for more information.
Reference
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Pedersen, Henrik, et al. "Structure-guided engineering of a complement component C3-binding nanobody improves specificity and adds cofactor activity." Frontiers in Immunology 13 (2022): 872536. Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only.
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