293F-spike–expressing cells were incubated with 100-fold diluted plasma samples for 30 min at 37°C. Cells were washed twice and stained with anti-human IgG PE, anti-human IgM Alexa Fluor 647, and anti-human IgA FITC (all from SouthernBiotech). Cells were then fixed with 4% formaldehyde solution, and fluorescence was evaluated on an LSR II (BD Biosciences).
Anti human iga fitc
Manufactured by Southern Biotech
Sourced in United States
Anti-human IgA FITC is a fluorescently labeled antibody that binds specifically to the IgA immunoglobulin found in human samples. It is a tool used in various immunoassay and flow cytometry applications to detect and quantify IgA levels.
Automatically generated - may contain errors
Lab products found in correlation
Anti human igg pe, by Southern Biotech (3 mentions)
Anti human igm alexa fluor 647, by Southern Biotech (3 mentions)
Formaldehyde solution, by Tousimis (1 mentions)
Freestyle 293 f, by Thermo Fisher Scientific (1 mentions)
Sars cov 2 spike protein, by GenScript (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Lsr 2 cytometer, by BD (1 mentions)
Anti iga biotin, by Southern Biotech (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Complete protease inhibitor, by Roche (1 mentions)
Streptavidin apc, by BD (1 mentions)
4 protocols using anti human iga fitc
1
SARS-CoV-2 Antibody Profiling Assay
2
SARS-CoV-2 Spike Protein Expression and Antibody Binding
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SARS-CoV-2 spike-expressing FreeStyle™ 293-F (RRID:CVCL_D603; Invitrogen cat. no. R79007) cells were generated by transfection with linearized plasmid (pcDNA3.1) encoding codon-optimized full-length SARS-CoV-2 spike protein (GenScript) matching the amino acid sequence having mutations E156G, D614G, P681R, T19R, T478K, L452R, D950N, and 157–158 del for Delta. Stable transfectants were single-cell sorted and selected to obtain a high-level Spike surface expressing clone (293F-Spike-Delta). 293F-Spike-Delta cells were incubated with 100 μl of 4-fold serial dilutions of plasma starting at 100-fold for 30 min at 4 °C. Cells were washed twice and stained with anti-human IgG PE, anti-human IgM Alexa Fluor 647, and anti-human IgA FITC (Southern Biotech, Birmingham, AL, USA, cat. nos. 2040-09, 2020-31, and 2050-02). Cells were then fixed with 4% formaldehyde solution (Tousimis, Rockville, MD, USA, cat. no. 1008B) and fluorescence was evaluated on a LSRII (BD Bioscience).
3
SARS-CoV-2 Spike Protein Binding Assay
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SARS-CoV-2 S-expressing expi293F cells were generated by transfection with linearized plasmid (pcDNA3.1) encoding codon-optimized full-length SARS-CoV-2 S protein matching the amino acid sequence of the IL-CDC-IL1/2020 isolate (GenBank accession number MN988713). Stable transfectants were single-cell sorted and selected to obtain a high-level S surface expressing clone (293F-spike-S2A). SARS-CoV-2 S-expressing cells were incubated with 200-fold diluted plasma samples for 30 min at 37 °C. Cells were washed twice and stained with anti-human IgG PE, anti-human IgM Alexa Fluor 647, and anti-human IgA FITC (Southern Biotech). Cells were then fixed with 4% formaldehyde solution, and fluorescence was evaluated on an LSRII flow cytometer (BD Bioscience).
4
Bacterial IgA Binding Assay
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Mouse fecal bacteria were isolated as described elsewhere 23 and were incubated for 1 hour on ice with sera, intestinal washes, or LNPC culture supernatant diluted in phosphate-buffered saline (PBS) þ protease inhibitor (cOmplete protease inhibitor, Roche, Meylan, France) to reach a concentration of 1 mg/mL of IgAs. After washings, bacteria were incubated with anti-IgA-biotin (Southern Biotech, Birmingham, AL) followed by streptavidin-APC (BD Pharmingen, Pont de Claix, France). Alternatively, ATCC strains of Escherichia coli (#8739), Enterococcus faecalis (#29212), Enterobacter cloacae (#13047) and Bacteroides vulgatus (#8482) were incubated (2.5 Â 10 7 bacteria) for 1 hour on ice with serial dilutions of human sera or human LNPC culture SN diluted in PBS þ protease inhibitor, washed, and further incubated with anti-human IgA-FITC (Southern Biotech). Bacterial suspensions then were fixed in 4% paraformaldehyde (PFA), washed, and suspended in PBS containing 2.5 mg/mL DAPI (Sigma Aldrich). Thirty thousand events were analyzed with the BD LSRII cytometer at a flow rate of 500-1500 events/s. Events that were DAPIpositive were considered as bacteria.
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