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Anti cd40 hm40 3

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Anti-CD40 (HM40-3) is a monoclonal antibody that binds to the CD40 receptor on the surface of B cells and other antigen-presenting cells. It is used as a research tool to study the role of CD40-mediated signaling in immune responses and cellular functions.

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Lab products found in correlation

Anti cd40, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Nacalai Tesque (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Celltrace violet dye, by Thermo Fisher Scientific (1 mentions) Cd43 microbead, by Miltenyi Biotec (1 mentions) Resiquimod r848, by InvivoGen (1 mentions)

Close spelling variants of this product

Anti-CD40 (HM40-3) agonistic antibody HM40-3 Anti-CD40

2 protocols using anti cd40 hm40 3

1

B Cell Activation Assay Protocol

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B cells were cultured in RPMI 1640 (Nacalai) supplemented with 10% FBS (Sigma-Aldrich), penicillin and streptomycin antibiotics (Nacalai), Hepes (Nacalai), glutamine (Nacalai), sodium pyruvate (Nacalai), nonessential amino acids (Nacalai), and β-mercaptoethanol (Nacalai). Purified naive B cells were stimulated with 1 µg/ml anti-CD40 (HM40-3; eBioscience) or 1 µg/ml F(ab′)2 goat anti-IgM (16-5092-85; eBioscience) or 1 µg/ml F(ab′)2 goat anti-IgG (16-5098-85; eBioscience). For ex vivo stimulation of Pre-GC, splenocytes from day 9 PyNL-infected mice were stimulated with 1 µg/ml anti-CD40 (eBioscience) and/or with 1 µg/ml anti-IgG (eBioscience) and 1 µg/ml anti-IgM (eBioscience). Cells were harvested at the time points indicated in the figure legends.
Lee M.S., Inoue T., Ise W., Matsuo-Dapaah J., Wing J.B., Temizoz B., Kobiyama K., Hayashi T., Patil A., Sakaguchi S., Simon A.K., Bezbradica J.S., Nagatoishi S., Tsumoto K., Inoue J.I., Akira S., Kurosaki T., Ishii K.J, & Coban C. (2021). B cell–intrinsic TBK1 is essential for germinal center formation during infection and vaccination in mice. The Journal of Experimental Medicine, 219(2), e20211336.
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2

B Cell Stimulation and Proliferation

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CD43– FO B cells were enriched from lymph nodes by immunomagnetic depletion of non-B cells with CD43-MicroBeads (Miltenyi Biotec). The sorted cells were labeled with 5 μM CellTrace™ Violet dye (Thermo Fisher Scientific) in PBS for 20 min at 37 °C. After washing with stimulation medium (RPMI-1640 supplemented with 10% heat-inactivated FCS, 1 mM glutamine and 50 μM β-mercaptoethanol), the cells were seeded at a density of 0.5 × 106 cells in 1 ml of stimulation medium containing 1.3 μg/ml anti-IgM antibody (II/41) plus 10 ng/ml IL-4 or 1.5 μg/ml anti-CD40 (HM40-3, eBioscience), 1 μg/ml HEL (Sigma/Roche), 1 μg/ml (0.2 μM) CpG oligodeoxynucleotides (ODN1826, InvivoGen), 2 μg/ml Pam3 (Pam3CSK4, Invivogen), 1 μg/ml Resiquimod (R848, Invivogen) or 25 μg/ml LPS. At day 4, the proliferation of the stimulated B cells was assessed by flow cytometric analysis. Live/dead separation for in vitro cultured cells was achieved by staining with fixable viability dye eFluor780 (Thermo Fisher Scientific).
Schwickert T.A., Tagoh H., Schindler K., Fischer M., Jaritz M, & Busslinger M. (2019). Ikaros prevents autoimmunity by controlling anergy and Toll-like receptor signaling in B cells. Nature immunology, 20(11), 1517-1529.
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