HEK293T cells were transfected with a plasmid-expressing wild-type SARS-CoV-2 S protein53 (link). Next day, transfected cells were collected and incubated with sera or recombinant anti-SARS-CoV-2 Spike Glycoprotein S1 antibody (CR3022, Abcam, Cat. No. ab273073, 1 µg/mL) for 30 min, washed twice with PBS/BSA and stained with anti-human IgG-Alexa647 (polyclonal, Southern Biotech Cat. No. 2014-31, 1:400), PE anti-IgM (CH2, Invitrogen Cat. No. MA1-10381, 1:200) and FITC anti-IgA (polyclonal, Southern Biotech Cat. No. 2052-02, 1:200), anti-human IgA1 Alexa488 (B3506B4, Southern Biotech Cat. No. 9130-30, 1:200), anti-human IgA2-Alexa647 (A9604D2, Southern Biotech Cat. No. 9140-31, 1:200) for 30 min. Cells were washed with PBS/BSA and DAPI was added before analysis for dead cell exclusion. Samples were analyzed on a FACSCanto (BD Biosciences) and using FlowJo v10 (Tree Star Inc.) analysis software. Mean fluorescent intensities were determined for transfected and non-transfected cells of each sample. Data were presented as ΔMFI = MFI(transfected)−MFI(non-transfected).
Ferreira-Gomes M., Kruglov A., Durek P., Heinrich F., Tizian C., Heinz G.A., Pascual-Reguant A., Du W., Mothes R., Fan C., Frischbutter S., Habenicht K., Budzinski L., Ninnemann J., Jani P.K., Guerra G.M., Lehmann K., Matz M., Ostendorf L., Heiberger L., Chang H.D., Bauherr S., Maurer M., Schönrich G., Raftery M., Kallinich T., Mall M.A., Angermair S., Treskatsch S., Dörner T., Corman V.M., Diefenbach A., Volk H.D., Elezkurtaj S., Winkler T.H., Dong J., Hauser A.E., Radbruch H., Witkowski M., Melchers F., Radbruch A, & Mashreghi M.F. (2021). SARS-CoV-2 in severe COVID-19 induces a TGF-β-dominated chronic immune response that does not target itself. Nature Communications, 12, 1961.
Alexa647 Anti iga Anti igg Anti igm AntibodyCellsCr3022Dapi Fitc HumanIga1 Iga2 Plasmid Sars cov 2 Sars cov 2 spike glycoprotein Sera Tree
Corresponding Organization :
Other organizations :
Leibniz Association, German Rheumatism Research Centre, Charité - Universitätsmedizin Berlin, Freie Universität Berlin, Humboldt-Universität zu Berlin, Friedrich-Alexander-Universität Erlangen-Nürnberg, Berlin Institute of Health at Charité - Universitätsmedizin Berlin
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
