Anti human igm alexa fluor 647

Manufactured by Southern Biotech

Sourced in United States

Anti-human IgM Alexa Fluor 647 is a fluorescently labeled antibody that binds to human immunoglobulin M (IgM). It is designed for use in various immunoassay and flow cytometry applications.

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Lab products found in correlation

Anti human igg pe, by Southern Biotech (3 mentions) Anti human iga fitc, 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)

3 protocols using anti human igm alexa fluor 647

SARS-CoV-2 Antibody Profiling Assay

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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).

Yu J., Thomas P.V., McMahan K., Jacob-Dolan C., Liu J., He X., Hope D., Martinez E.J., Chen W.H., Sciacca M., Hachmann N.P., Lifton M., Miller J., Powers O.C., Hall K., Wu C., Barrett J., Swafford I., Currier J.R., King J., Corbitt C., Chang W.C., Golub E., Rees P.A., Peterson C.E., Hajduczki A., Hussin E., Lange C., Gong H., Matyas G.R., Rao M., Paquin-Proulx D., Gromowski G.D., Lewis M.G., Andersen H., Davis-Gardner M., Suthar M.S., Michael N.L., Bolton D.L., Joyce M.G., Modjarrad K, & Barouch D.H. (2022). Protection against SARS-CoV-2 Omicron BA.1 variant challenge in macaques by prime-boost vaccination with Ad26.COV2.S and SpFN. Science Advances, 8(47), eade4433.

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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).

Gromowski G.D., Cincotta C.M., Mayer S., King J., Swafford I., McCracken M.K., Coleman D., Enoch J., Storme C., Darden J., Peel S., Epperson D., McKee K., Currier J.R., Okulicz J., Paquin-Proulx D., Cowden J, & Peachman K. (2023). Humoral immune responses associated with control of SARS-CoV-2 breakthrough infections in a vaccinated US military population. eBioMedicine, 94, 104683.

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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).

King H.A., Joyce M.G., Lakhal-Naouar I., Ahmed A., Cincotta C.M., Subra C., Peachman K.K., Hack H.R., Chen R.E., Thomas P.V., Chen W.H., Sankhala R.S., Hajduczki A., Martinez E.J., Peterson C.E., Chang W.C., Choe M., Smith C., Headley J.A., Elyard H.A., Cook A., Anderson A., Wuertz K.M., Dong M., Swafford I., Case J.B., Currier J.R., Lal K.G., Amare M.F., Dussupt V., Molnar S., Daye S.P., Zeng X., Barkei E.K., Alfson K., Staples H.M., Carrion R., Krebs S.J., Paquin-Proulx D., Karasavvas N., Polonis V.R., Jagodzinski L.L., Vasan S., Scott P.T., Huang Y., Nair M.S., Ho D.D., de Val N., Diamond M.S., Lewis M.G., Rao M., Matyas G.R., Gromowski G.D., Peel S.A., Michael N.L., Modjarrad K, & Bolton D.L. (2021). Efficacy and breadth of adjuvanted SARS-CoV-2 receptor-binding domain nanoparticle vaccine in macaques. Proceedings of the National Academy of Sciences of the United States of America, 118(38), e2106433118.

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