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Igm fitc

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IgM FITC is a fluorescently labeled antibody that specifically binds to the IgM immunoglobulin. It can be used to detect and quantify IgM molecules in various applications, such as flow cytometry and immunoassays.

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Lab products found in correlation

Ifn γ apc, by BioLegend (1 mentions) Cd4 fitc, by BioLegend (1 mentions) Igg1 pe, by BioLegend (1 mentions) Anti fcγrii fcγriii 2.4g2, by BD (1 mentions) Il 2 apc, by BioLegend (1 mentions) Plus brefeldin a, by Thermo Fisher Scientific (1 mentions) Cd138 apc, by BioLegend (1 mentions) Cd5 pe cy7, by BioLegend (1 mentions) B220 fitc, by BioLegend (1 mentions) Il 17 pe, by Miltenyi Biotec (1 mentions) Cd1d pe, by BioLegend (1 mentions) Facscanto 2, by BD (1 mentions) Anti cd16 cd32, by BD (1 mentions) Igd apc, by BioLegend (1 mentions) Cd38 pe, by BioLegend (1 mentions) Facs attune, by Thermo Fisher Scientific (1 mentions) Anti mouse b220 pe cy7, by BioLegend (1 mentions) Igd apc cy7, by BioLegend (1 mentions) Cd44 fitc, by BioLegend (1 mentions) Cd19 pacific blue, by BioLegend (1 mentions)

Close spelling variants of this product

Anti-IgM FITC FITC anti-mouse IgM Anti-human IgM-FITC FITC Mouse IgM k Isotype

4 protocols using igm fitc

1

B Cell Immunophenotyping and Cytokine Analysis

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B cells were washed once in cold PBS containing 0.1% BSA (FACS buffer) before blocking with anti-FcγRII/FcγRIII (2.4G2, BD Pharmingen, San Diego, CA). Stainings were performed on ice using conjugated mAbs (eBioscience, San Diego, CA, if not stated otherwise), diluted 1:300 in FACS buffer for surface marker and 1:200 for intracellular cytokine staining followed by incubation for 20 min. After washing with FACS buffer, cells were analysed on a FACS Canto II (BD) using FlowJo software (Tree star, Ashland, OR). The following Abs were used: B220-FITC (#11-0452-86), CD5-PE-Cy7 (#25-0051-81), CD1d-PE (#12-0011-81), CD138-APC (#142506, Biolegend), IgM-FITC (#11-5890-85), IgG1-PE (#12-4015-82), CD4-FITC (#11-0041-82), IL-2-APC (#17-7021-81), IFNγ-APC (#17-7311-82) and IL-10-PerCP (#45-7101-80). Abs against TNF−α−PE (#130-092-245) and IL-17-PE (#130-094-296) were from Miltenyi Biotec. For intracellular staining, the fixation and permeabilization kit (Plus Brefeldin A; eBioscience, Cat. no. 88-8823-88) was used according to manufacturer's recommendation.
Alrefai H., Muhammad K., Rudolf R., Pham D.A., Klein-Hessling S., Patra A.K., Avots A., Bukur V., Sahin U., Tenzer S., Goebeler M., Kerstan A, & Serfling E. (2016). NFATc1 supports imiquimod-induced skin inflammation by suppressing IL-10 synthesis in B cells. Nature Communications, 7, 11724.
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2

Profiling Memory B Cells from Vaccination

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To quantify and profile memory B cells from vaccination, single‐cell suspensions were prepared from inguinal lymph nodes, as previously described.32 Cells were stained with Zombie live/dead (Biolegend, San Diego, CA, USA), FcR‐blocked (anti‐CD16/CD32, BD Bioscience, Franklin Lakes, NJ, USA) and stained with a cocktail containing anti‐mouse B220‐PECy7, CD38‐PE, IgD‐APC, CD95 (Fas)‐BV605, IgM‐FITC, IgG‐APCCy7 (all Biolegend), for 30 min on ice in FACS buffer (PBS, 1% FBS, 0.5% NaN3). Cells were finally fixed with 100 µL of 4% PFA for 20 min on ice. B memory cells were defined as B220+ CD38+/− Faslow IgD‐ IgM+ or IgG+. Samples were acquired by flow cytometry on a FACS Attune (Invitrogen) and analysed with FlowJo software (BD). Representative FACS plots are shown in Supplementary figure 2a.
Kavian N., Hachim A., Cowling B.J, & Valkenburg S.A. (2021). Repeated influenza vaccination provides cumulative protection from distinct H3N2 viruses. Clinical & Translational Immunology, 10(6), e1297.
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3

Immunophenotyping B and T Cells

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WT and mFICD−/− mice aged 6–8 weeks were euthanized by CO2 asphyxiation followed by cervical dislocation. Spleen, bone marrow, and thymus tissues were extracted and homogenized in PBE buffer (PBS +0.5% BSA and 1 mM EDTA). Red blood cells were lysed using ACK lysis buffer (Gibco), and cells were resuspended in PBE for surface staining with the following antibodies (clone;source) for 30 min at 4˚C: B220 PerCP-Cy5.5, CD43 APC, CD19 Pacific Blue, IgM FITC, IgD APC-Cy7, CD21 PerCP-Cy5.5, CD23 FITC, IgM APC, CD44 FITC, CD25 PE-Cy7, CD4 APC, CD8α Pacific Blue (53–6.7;BioLegend), CD3 PerCP-Cy5.5 (145-2C11;BD), B220 APC-Cy7. All samples were blocked using Fc-Block (BD Bioscience). Acquisition of B and T cell populations was performed on an LSRFortessa cytometer (BD) instrument and analyzed with the FlowJo software package (Tree-Star).
McCaul N., Porter C.M., Becker A., Tang C.H., Wijne C., Chatterjee B., Bousbaine D., Bilate A., Hu C.C., Ploegh H, & Truttmann M.C. (2021). Deletion of mFICD AMPylase alters cytokine secretion and affects visual short-term learning in vivo. The Journal of Biological Chemistry, 297(3), 100991.
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4

Characterization of Lymph Node Immune Cells

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Mandibular lymph nodes were macerated over 40-μm mesh cell strainer (Midsci, Valley Park, MO) into single-cell suspensions. Cells were enumerated and 1 × 106 cells/100 μl PBS containing 2% FBS were blocked with anti-CD16/32 (eBioscience, San Diego, CA), labeled with a combination of 1 μl each of CD45 PerCP-Cy5.5 (Biolegend, San Diego, CA), CD19 APC, CD3e, CD4 APC-Cy7, CD8a PE or APC-Cy7, IgM FITC, IgD PE-Cy7, CD44 APC, and/or CD62L FITC (all from Thermo Fischer Scientific) diluted in 1% BSA in 1X PBS for 30 minutes. In the case of tetramer staining, cells were labeled with a combination of CD3 PE-Cy7, CD8-APC-Cy7, gB (SSIEFARL)-PE or ICP6 (QTFDFGRL)-Alexafluor488 (NIH Tetramer Core Facility, Atlanta, GA). Cells were then washed twice by adding 1 ml of 2% FBS in 1X PBS, centrifuging for 5 minutes at 300 × g, and decanting supernatant. Cells were then fixed in 1 ml of 1% paraformaldehyde overnight and resuspended in 1 ml of 2% FBS in 1X PBS to be analyzed on a MacsQuant 196 flow cytometer (Miltenyi Biotech). Gating strategies were identical to those previously described [40 (link)] except those included in this study, incubated on ice in the dark for 20–30 min, and washed in PBS containing 2% FBS. Samples were analyzed using FlowJo software (Ashland, OR).
Carr D.J., Berube A.N., Filiberti A, & Gmyrek G.B. (2021). Lack of Neonatal Fc Receptor does not Diminish the Efficacy of the HSV-1 0ΔNLS Vaccine Against Ocular HSV-1 Challenge. Vaccine, 39(18), 2526-2536.
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