Creative Biolabs is a global company with extensive scientific expertise in the field of complement. Based on years of experience, now we are confident in offering high-quality products and the best service for our clients all over the world. Here, we describe the recombinant C2 reverse-phase analysis protocol to promote your research. Please note that our protocols are only for your reference.
Recombinant C2 Reverse-Phase Analysis
Fig.1 Flow chart of recombinant C2 reverse-phase analysis. (Creative Biolabs)
Published Data
Fig.2 Preparative workflow for C2 and C4b2 samples intended for small angle x-ray scattering characterization.1
To investigate the solution structure of C2, recombinant protein was produced via expression in HEK293 cells. Deglycosylated C2 was obtained by incubating the protein with a mixture of four distinct endoglycosidases designed to remove as many glycans as possible. Importantly, both native and deglycosylated C2 maintained binding to C4b. Given that C2 harbors eight Asn-linked glycans, each contributing approximately 2 kDa, the additional mass of 16 kDa is consistent with an 83 kDa protein sequence and an observed ~100 kDa band on SDS-PAGE. Moreover, size exclusion chromatography was employed to analyze purified C4b, C2, and the C4b2 complex formed upon their mixture.
As a leading products and custom services provider in the United State, Creative Biolabs now offers a range of complementary products and complement assay services through our established platform and protocols. For example, we can provide therapeutic antibodies, serine protease inhibitors, soluble complement regulators, complement component inhibitors, anaphylatoxin receptor antagonists, and other small molecules.
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Reference
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Mortensen, Sofia, Jan K. Jensen, and Gregers R. Andersen. "Solution structures of complement C2 and its C4 complexes propose pathway-specific mechanisms for control and activation of the complement proconvertases." Journal of Biological Chemistry 291.32 (2016): 16494-16507. Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only.
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