Complement Deposition Assay Protocol

Complement deposition was performed as described before (45 (link)). In brief, biotinylated antigens were coupled to fluorescent neutravidin beads (Thermo Fisher) and incubated with 10 μl of 1:10 diluted plasma. Beads were incubated with guinea pig complement in GVB++ buffer (Boston BioProducts, MA, USA) for 20 min at 37°C. Deposited C3 on beads was stained with anti-guinea pig C3 antibody labeled with fluorescein isothiocyanate (FITC) (MP Biomedicals, CA, USA) and analyzed by flow cytometry on a LSRII instrument (BD Biosciences, CA, USA).

Free full text: Click here

Bartsch Y.C., Loos C., Rossignol E., Fajnzylber J.M., Yuan D., Avihingsanon A., Ubolyam S., Jupimai T., Hirschel B., Ananworanich J., Lauffenburger D.A., Li J.Z., Alter G, & Julg B. (2021). Viral Rebound Kinetics Correlate with Distinct HIV Antibody Features. mBio, 12(2), e00170-21.

Publication 2021

fluorescent neutravidin beads GVB++ buffer FITC LSRII instrument

Antibody Antigens Buffer Flow cytometry Fluorescein Guinea pig Isothiocyanate Neutravidin Plasma

Corresponding Organization : Ragon Institute of MGH, MIT and Harvard

Other organizations : Massachusetts Institute of Technology, Brigham and Women's Hospital, Harvard University, HIV Netherlands Australia Thailand Research Collaboration, Chulalongkorn University, University Hospital of Geneva, University of Amsterdam

Top 5 similar protocols

Variable analysis

independent variables

Biotinylated antigens coupled to fluorescent neutravidin beads
Plasma dilution (1:10)

dependent variables

Deposited C3 on beads
C3 staining with anti-guinea pig C3 antibody labeled with FITC

control variables

Guinea pig complement in GVB++ buffer
Incubation time (20 min)
Incubation temperature (37°C)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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!