BCR-Abl+ B-ALL and Top Notch T-ALL cells were cultured in RPMI-1640 (HyClone) with 10% fetal bovine serum (FBS), 5μM β-mercaptoethanol, and 4mM L-glutamine. Eu-myc Arf-/- cells were cultured in 45% DMEM/45% IMDM (HyClone) with 10% FBS, 5μM β-mercaptoethanol, and 2mM L-glutamine. K-rasLA2/+; p53LSL/LSL T cell lymphoma cells were cultured in IMDM with 10% FBS and 10μM β-mercaptoethanol. Pre-B cells were harvested from bone marrow of a C57BL/6 mouse, stained with fluorescently-conjugated anti-B220 (BioLegend), anti-CD11b (eBioscience), and anti-IgM antibodies (eBioscience) and sorted to obtain B220+CD11b-IgM- cells. Pre-B cells were cultured in 45% DMEM/45% IMDM with 10% FBS, 5μM β-mercaptoethanol, 2mM L-glutamine, recombinant murine IL-7 (1.0 ng/mL), and recombinant murine SCF (1.0 ng/mL) (Peprotech) on a feeder layer of bone marrow stromal cells.
Anti igm antibody
Manufactured by Thermo Fisher Scientific
Sourced in Germany
Anti-IgM antibodies are laboratory reagents used to detect and measure the presence of immunoglobulin M (IgM) in biological samples. IgM is an antibody class that is typically produced during the early stages of an immune response. These antibodies can be used in various immunoassay techniques to identify and quantify IgM levels, which can provide information about the immune status or disease progression in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd11b, by Thermo Fisher Scientific (2 mentions)
Anti b220, by BioLegend (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Recombinant murine il 7, by Thermo Fisher Scientific (1 mentions)
Recombinant murine scf, by Thermo Fisher Scientific (1 mentions)
Cd11b, by Thermo Fisher Scientific (1 mentions)
Anti f4 80, by Thermo Fisher Scientific (1 mentions)
Anti nk1, by Thermo Fisher Scientific (1 mentions)
Anti ly6g, by Thermo Fisher Scientific (1 mentions)
Anti cd8, by Thermo Fisher Scientific (1 mentions)
Anti cd11c, by Thermo Fisher Scientific (1 mentions)
Anti ly6c, by Thermo Fisher Scientific (1 mentions)
Anti cd19, by Thermo Fisher Scientific (1 mentions)
Anti mhcii, by Thermo Fisher Scientific (1 mentions)
Anti cd115, by Thermo Fisher Scientific (1 mentions)
Anti cd5, by Thermo Fisher Scientific (1 mentions)
Calibrite, by BD (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Anti cd23, by Thermo Fisher Scientific (1 mentions)
Anti cd21, by Thermo Fisher Scientific (1 mentions)
4 protocols using anti igm antibody
1
Culturing Murine Leukemia/Lymphoma Cell Lines
2
Intracellular Cytokine Staining of Immune Cells
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For intracellular-cytokine staining, single-cell suspended splenocytes were incubated with brefeldin A (eBioscience), followed by an additional 5 h of incubation at 37°C. After surface staining with anti-CD3, anti-CD8, anti-CD11b, anti-CD11c, anti-CD19, anti-CD115, anti-F4/80, anti-Ly6C, anti-Ly6G, anti-MHC-II, and anti-NK1.1 antibodies (all from eBioscience), the cells were fixed with 2% formalin, permeabilized with 0.1% saponin, and stained with anti-TNF antibodies (eBioscience) for 30 min at 4°C. B-cell subsets were detected in single-cell suspensions of splenocytes with anti-CD5, anti-CD19, anti-CD21, anti-CD23, and anti-IgM antibodies (all from eBioscience). BD Calibrite (BD Biosciences, USA) beads were added to the samples before acquisition with a BD LSRFortessa.
3
Immunomodulatory Treatments and Cell Responses
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In some cases, cell lines were stimulated for 48 hrs prior with treatment of drugs (RTX or obinutuzumab or silencing with PLCG2), with or without anti-IgM antibody (50 ug/ml, Invitrogen, Grand Island, NY). Stimulated cells were used for cell proliferation or apoptosis assay as described above.
4
Immunofluorescence Analysis of Renal C4d, C3d, and IgM
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Renal C4d deposition was investigated using a specific antibody for immunofluorescence [28 (link)], kindly provided by Dr. W. M. Baldwin III, and an antibody directed against C4d from Hycult Biotech (HP8034; Uden, The Netherlands). C3d deposition was analysed using an anti-C3d antibody (AF2655, R&D Systems, Abingdon, UK). Deposited IgM was detected using an FITC-labelled anti-IgM antibody (# 11-0990-82, Invitrogen, Darmstadt, Germany). Cryosections about 5 µm thick were made from the grafts fixed in melting 2-methylbutane. Cryosections were incubated for 1 h at room temperature in PBS containing 10% normal goat serum (pH 7.4) and then for 90 min at room temperature with affinity-purified primary antibodies at a dilution of 1/200 (kidney sections). After washing in PBS, the sections were incubated for 45 min at room temperature with PBS-diluted secondary antibodies (Alexa Fluor 488 goat anti-rabbit-Ig, 1/1000; Invitrogen, Karlsruhe, Germany). Sections were rinsed with PBS, coverslipped with Crystal Mount (GeneTex, San Antonio, TX, USA), and evaluated by epifluorescence microscopy (Observer Z1 with apotome; Zeiss, Göttingen, Germany).
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