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Igm clone mhm 88

Manufactured by BioLegend
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IgM (clone MHM-88) is a monoclonal antibody product manufactured by BioLegend. This antibody specifically recognizes the IgM immunoglobulin.

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Lab products found in correlation

Cd19 clone hib19, by Thermo Fisher Scientific (2 mentions) Cd27 clone o323, by BioLegend (2 mentions) Igd clone ia6 2, by BioLegend (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Aria 2, by BD (1 mentions) Viability stain, by Thermo Fisher Scientific (1 mentions) Lsr fortessa cytometer, by BD (1 mentions) Fc blocked, by BD (1 mentions) Cd38 clone hit2, by BioLegend (1 mentions) Macsquant 16 flow cytometer, by Miltenyi Biotec (1 mentions) Cd27 clone m t271, by BioLegend (1 mentions) Cd3 clone okt3, by BioLegend (1 mentions) Cd95 clone dx2, by BioLegend (1 mentions) Cd14 clone m5e2, by BioLegend (1 mentions) Cd8 clone rpa t8, by BD (1 mentions) Cd3 clone sp34 2, by BD (1 mentions) Cd4 clone okt4, by BioLegend (1 mentions) Cd20 clone 2h7, by BioLegend (1 mentions) Igg clone g18 145, by BD (1 mentions) Cd14 m5e2, by BD (1 mentions)

Close spelling variants of this product

MHM-88 (7 citations) IgM (MHM-88, (6 citations) ), APC anti-IgM (clone MHM-88, (2 citations)

4 protocols using igm clone mhm 88

1

Sorting Naive and Memory B Cells

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PBMCs were stained with antibodies specific for CD19 (clone HIB19, eBiosciences, San Diego, CA), IgM (clone MHM-88), IgD (clone IA6-2), and CD27 (clone O323, all from BioLegend, San Diego, CA). B cell populations were sorted using an Aria II (BD Biosciences) into naive (CD19+CD27- and IgM+ and/or IgD+) or memory (CD19+CD27+) cells. Purity of sorted populations was tested and confirmed to have <2% contamination by the other B cell subset. Cells were lysed using TRIzol Reagent (Life Technologies, Grand Island, NY) and stored at -80°C until RNA extraction.
Roskin K.M., Simchoni N., Liu Y., Lee J.Y., Seo K., Hoh R.A., Pham T., Park J.H., Furman D., Dekker C.L., Davis M.M., James J.A., Nadeau K.C., Cunningham-Rundles C, & Boyd S.D. (2015). IgH sequences in common variable immune deficiency reveal altered B cell development and selection. Science translational medicine, 7(302), 302ra135.
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2

Multiparametric Flow Cytometry Analysis of Stimulated B Cells

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B cells were Fc blocked (BD Biosciences, San Jose, CA) prior to staining. Cells were analyzed using an LSR Fortessa cytometer (BD Biosciences), with gating performed by first setting a wide lymphocyte gate by forward and side scatter, followed by two sets of doublet exclusion, first by FSC-A by FSC-H and then by FSC-A by FSC-W, and by excluding dead cells with a viability stain (Life Technologies, Grand Island, NY). B cell purity was determined during isolation and at baseline by staining for CD19 (clone HIB19, eBiosciences, San Diego, CA). Cell phenotype was determined by staining IgM (clone MHM-88), CD27 (clone O323), and CD38 (clone HIT2, all from BioLegend, San Diego, CA), with results reported for the live cells gate. B cell proliferation was determined by dilution of CFSE dye, with results reported for the live singlet lymphocyte gate.
Simchoni N, & Cunningham-Rundles C. (2015). TLR7 and TLR9 responsive human B cells share phenotypic and genetic characteristics. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3035-3044.
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3

Characterizing B cell subsets by flow cytometry

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After preparation of freshly isolated PBMCs in flow cytometry buffer, PBMCs were stained with fluorescent labeled antibodies and measured on a MACSQuant 16 flow cytometer (Miltenyi Biotec, Bergisch Gladbach, Germany). B cell subsets were stained using antibodies against CD19 (clone REA675, Miltenyi), CD20 (clone REA780, Miltenyi), CD27 (clone M-T271, Biolegend, San Diego, California, US), CD138 (clone BB4/MI15, Biolegend), HLA-DR (clone REA332, Miltenyi), IgD (clone REA740, Miltenyi), IgM (clone MHM-88, Biolegend), IgA (clone M24A, Merck Millipore), CD95 (clone DX2, Biolegend) as well as CD3 (clone OKT3, Biolegend) and CD14 (clone M5E2, Biolegend) for the dump channel. Definition of B cell populations: naive B cells: CD19+CD20+CD27-; memory B cells: CD19+CD20+CD27+; Plasmablasts and plasma cells: CD19+CD20-CD27hi).
Bernardes J.P., Mishra N., Tran F., Bahmer T., Best L., Blase J.I., Bordoni D., Franzenburg J., Geisen U., Josephs-Spaulding J., Köhler P., Künstner A., Rosati E., Aschenbrenner A.C., Bacher P., Baran N., Boysen T., Brandt B., Bruse N., Dörr J., Dräger A., Elke G., Ellinghaus D., Fischer J., Forster M., Franke A., Franzenburg S., Frey N., Friedrichs A., Fuß J., Glück A., Hamm J., Hinrichsen F., Hoeppner M.P., Imm S., Junker R., Kaiser S., Kan Y.H., Knoll R., Lange C., Laue G., Lier C., Lindner M., Marinos G., Markewitz R., Nattermann J., Noth R., Pickkers P., Rabe K.F., Renz A., Röcken C., Rupp J., Schaffarzyk A., Scheffold A., Schulte-Schrepping J., Schunk D., Skowasch D., Ulas T., Wandinger K.P., Wittig M., Zimmermann J., Busch H., Hoyer B.F., Kaleta C., Heyckendorf J., Kox M., Rybniker J., Schreiber S., Schultze J.L, & Rosenstiel P. (2020). Longitudinal Multi-omics Analyses Identify Responses of Megakaryocytes, Erythroid Cells, and Plasmablasts as Hallmarks of Severe COVID-19. Immunity, 53(6), 1296-1314.e9.
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4

Isolation and Screening of Antigen-Specific Memory B Cells

Cited 1 time
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Single cell sorting was performed as previously described (Mason et al., 2016 (link)). In brief, rhesus PBMCs were stained with an antibody cocktail of CD3 (clone SP34–2, BD Biosciences), CD4 (clone OKT4, BioLegend), CD8 (clone RPA-T8, BD Biosciences), CD14 (M5E2, BD Biosciences), CD20 (clone 2H7, Biolegend), IgM (clone MHM-88, Biolegend), IgG (clone G18–145, BD Biosciences) and BG505 SOSIP-AviB conjugated with streptavidin-Alexa Fluor 647 (AF647) and streptavidin-Alexa Fluor 488 (AF488) respectively. Cells were stained at room temperature for 20 mins in the dark. Antigen-specific memory B cells (CD3−CD4−CD8−CD14−CD20+IgM−IgG+BG505 SOSIP.664 dual positive) were single-cell sorted into 384-well culture plates according to the published protocol (Huang et al., 2013 (link)). After 2 weeks of culture, supernatants were harvested and screened for neutralization activity. Ig genes from neutralization positive wells were amplified by RT-PCR, nested PCR, then cloned into expression vectors and expressed in 293F cells.
Nogal B., Bianchi M., Cottrell C.A., Kirchdoerfer R.N., Sewall L.M., Turner H.L., Zhao F., Sok D., Burton D.R., Hangartner L, & Ward A.B. (2020). Mapping Polyclonal Antibody Responses in Non-human Primates Vaccinated with HIV Env Trimer Subunit Vaccines. Cell reports, 30(11), 3755-3765.e7.
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