Purification and Quantification of C1q and Related Proteins

C1q was purified from human serum and quantified as described (12 (link)). Human serum was obtained from the Etablissement Français du Sang (EFS) Rhône-Alpes (agreement number 14-1940 regarding its use in research). C1q collagen stalks (CLR) and C1q globular heads (GR) were prepared according to Tacnet-Delorme et al. (13 (link)). The recombinant protease tetramer C1r2s2 was produced and purified according to Bally et al. (14 (link)). Full-length LRP1 (soluble LRP1) was produced and purified according to De Nardis et al. (15 (link)). Recombinant C1q and C1q mutant LysA59Ala/LysB61Ala/LysC58Ala were produced and purified as described in Bally et al. (16 (link)). For protein quantification, Mw and A_{1%, 1 cm} were respectively for C1q (459,300; 6.8) (12 (link)), CLR (189,900; 2.1), GR (48,000; 9.3) (17 (link)), C1r2s2 (330,000; 13.5) (18 (link)). Oligonucleotides were from Eurogentec. Restriction and modification enzymes were from New England Biolabs.
Full-length LRP1 Myc DDK clone (RC218369) was purchased from Origene. The pET22B-RAP plasmid was kindly provided by Søren Moestrup, Aahrus University, Denmark. pcDNA3.1 mini IV HA-tag was cloned as previously indicated (19 (link)).

Free full text: Click here

Fouët G., Gout E., Wicker-Planquart C., Bally I., De Nardis C., Dedieu S., Chouquet A., Gaboriaud C., Thielens N.M., Kleman J.P, & Rossi V. (2020). Complement C1q Interacts With LRP1 Clusters II and IV Through a Site Close but Different From the Binding Site of Its C1r and C1s-Associated Proteases. Frontiers in Immunology, 11, 583754.

Publication 2020

Oligonucleotides Restriction and modification enzymes

Collagen Enzymes Globular Heads Human Oligonucleotides Plasmid Protease Protein Serum Stalks Tetramer

Corresponding Organization :

Other organizations : Institut de Biologie Structurale

Top 5 similar protocols

Variable analysis

independent variables

Purified C1q from human serum
Prepared C1q collagen stalks (CLR) and C1q globular heads (GR)
Produced and purified recombinant protease tetramer C1r2s2
Produced and purified full-length LRP1 (soluble LRP1)
Produced and purified recombinant C1q and C1q mutant LysA59Ala/LysB61Ala/LysC58Ala

dependent variables

Quantification of purified C1q from human serum

control variables

Human serum obtained from Etablissement Français du Sang (EFS) Rhône-Alpes
Molecular weights and extinction coefficients used for protein quantification

positive controls

None specified

negative controls

None specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!