αcd40

Manufactured by BioXCell

Sourced in United States

αCD40 is a monoclonal antibody that binds to the CD40 receptor on the surface of antigen-presenting cells, such as B cells, dendritic cells, and macrophages. The CD40 receptor is involved in the activation and regulation of the immune response.

Automatically generated - may contain errors

Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Poly 1 c, by Cytiva (1 mentions) Mog35 55, by Peptide Institute (1 mentions) α actin, by Merck Group (1 mentions) Apc cy7, by BioLegend (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Tgf β, by R&D Systems (1 mentions) Rmpi 1640, by Thermo Fisher Scientific (1 mentions) Recombinant mouse il 10, by BioLegend (1 mentions) Rat igg2a, by BioXCell (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Recombinant murine il 21, by Thermo Fisher Scientific (1 mentions) α4 1bb, by BioXCell (1 mentions)

7 protocols using αcd40

Investigating gene expression and CD11b after CD40 stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

To investigate gene expression after αCD40 stimulation, murine splenic B cells were plated at 2.5 × 10⁶/ml in 24-well plates in B cell medium: RMPI 1640 + 10% FBS (Life Technologies, Carlsbad, CA, USA) + penicillin-streptomycin (100 U/ml, Thermo Fisher Scientific, Waltham, MA, USA). Cells were incubated with 10 μg/ml of αCD40 (clone: FGK4.5) (BioXCell, Lebanon, NH, USA) or rat IgG2a (clone: 2A3, BioXCell) for 6, 24, 48 h, and collected for gene expression analysis.
To investigate whether CD40 stimulation or IL-10 affected CD11b expression, murine splenic B cells were plated at 2.0 × 10⁶ cells/ml in B cell medium + 5 mM of Mg²⁺ in 96-well plates. Cells were incubated for 48 h with (a) medium alone, (b) 2 µg/ml of lipopolysaccharide (LPS, Sigma-Aldrich, St. Louis, MO, USA), (c) 2 µg/ml of LPS + 10 μg/ml of rat IgG2a (clone: 2A3, BioXCell), (d) 2 µg/ml of LPS + 10 μg/ml of αCD40 (clone: FGK4.5, BioXCell), (e) 2 µg/ml of LPS + 50 ng/ml of IL-10 (Recombinant mouse IL-10, Biolegend, San Diego, CA, USA). After 48 h, cells were collected and stained to assess CD11b surface expression by flow cytometry.

van Hooren L., Vaccaro A., Ramachandran M., Vazaios K., Libard S., van de Walle T., Georganaki M., Huang H., Pietilä I., Lau J., Ulvmar M.H., Karlsson M.C., Zetterling M., Mangsbo S.M., Jakola A.S., Olsson Bontell T., Smits A., Essand M, & Dimberg A. (2021). Agonistic CD40 therapy induces tertiary lymphoid structures but impairs responses to checkpoint blockade in glioma. Nature Communications, 12, 4127.

+ Open protocol

+ Expand

Immune Response Induction by Peptides

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunizations consisted of either100 μg B8R_20-27 (TSYKFESV), 100 μg gp33 (KAVYNFATM) (both provided by S. Jameson), 200 μg MOG_35-55 (MEVGWYRSPFSRVVHLYRNGK) (Peptides International), or 50 μg PLP_178-191 (NTWTTCQSIAFPSK) (Peptides International) mixed with 50 μg of poly I:C (Amersham) and 50 μg of α-CD40 (BioXcell), or 100 μg 2W1S (EAWGALANWAVDSA) (GenScript Corp.) mixed with 5 μg LPS (List Biologicals) diluted in 1x PBS to a total volume of 100 μL per injection. Injections were administered retro-orbitally under isoflurane anesthesia.

Schuldt N.J., Auger J.L., Hogquist K.A, & Binstadt B.A. (2015). Bi-Allelic TCRα or β Recombination Enhances T Cell Development but Is Dispensable for Antigen Responses and Experimental Autoimmune Encephalomyelitis. PLoS ONE, 10(12), e0145762.

+ Open protocol

+ Expand

B cell NF-κB activation protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

B cells were isolated by magnetic bead isolation with mouse Pan B Cell Isolation beads (Miltenyi Biotec, #130-095-813) using the manufacturers provided protocol. Isolated B cells were exposed to 10 µg/ml αCD40 (BioXCell, #BE0016-2) for 4 h. Proteins were measured by western blot by staining with αTNIK (Abcam, ab95887), αNF-κB p65 (cell Signaling, #3033) and αPhospho-NF-κB p65 (Cell signaling, #6956). αAlpha-tubulin (Sanbio, CLT9002) or αActin (Sigma-Aldrich A3853) were used as a loading control. TNIK and NF-κB p65 were visualized by staining with HRP conjugated Goat anti-Rabbit (Invitrogen, #32260) and Goat anti-Mouse (Invitrogen, #32330). Phospho-NF-κB p65 and alpha-tubulin were staining with IRDye 800CW conjugated Goat anti-Mouse (Odyssey, #926-32210) and Goat anti-Rabbit IRDye 680RD (Odyssey, #926-68071). Data was analyzed using FiJi.

van Os B.W., Kusters P.J., den Toom M., Beckers L., van Tiel C.M., Vos W.G., de Jong E., Kieser A., van Roomen C., Binder C.J., Reiche M.E., de Winther M.P., Bosmans L.A, & Lutgens E. (2023). Deficiency of germinal center kinase TRAF2 and NCK-interacting kinase (TNIK) in B cells does not affect atherosclerosis. Frontiers in Cardiovascular Medicine, 10, 1171764.

+ Open protocol

+ Expand

Isolation and Differentiation of Plasma Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naive B cells were isolated from Peyer's patches and spleen using magnetic CD43⁺ bead isolation (Miltenyi Biotec, #130-049-801) following the manufacturer's provided protocol. To generate IgG producing plasma cells, naive B cells were exposed to 50 µg/ml LPS (InvivoGen, tlrl-pb5lps) and 10 ng/ml IL-4 (Peprotech, #214-14) for 3 days. To generate IgA producing plasma cells, naive B cells were exposed to 20 μg/ml αIgD (ThermoFisher scientific, 16-5924), 5 ng/ml IL-4 (Peprotech, #214-14), 1 ng/ml IL-5 (R&D systems, 405-ML), 10 ng/ml TGF-β (R&D systems, 7666-MB) and 20 μg/ml αCD40 (BioXcell, BE0016-2). Cells were stained in FACS buffer (0.5% bovine serum albumin, 5 mM EDTA in PBS; pH 7.4) with the following antibodies: αCD3 (1:100, APC, Biolegend, #100312), αCD11b (1:100, APC, BD Biosciences, #553312), αCD45 (1:100, APC-Cy7, Biolegend, #103115), αIgA (1:100, PE, ThermoFisher scientific, #12-4201), αIgG (1:200, FITC, Biolegend, #406001), αIgM (1:1,000, PE-Cy7, ThermoFisher scientific, #25-5790).

+ Open protocol

+ Expand

Modulating Splenic B Cell Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total splenocytes were isolated from B1-8i C57BL/6 mice in Stem Cell Buffer, ACK lysed for 60 seconds, washed once in Stem Cell Buffer then twice with PBS, and 2×10⁷ cells were injected into the tail veins of IL-21RKO Rag2KO C57BL/6 mice. Mice were immunized with NP-CGG the following day. At day 10 post-immunization, PBS, 5 μg recombinant murine IL-21 (PeproTech) and/or 50 μg α-CD40 (BioXCell) were injected into the tail veins as indicated, and euthanized for FACS staining 30 minutes (pSTAT3) or 4 hours (BCL6/IRF4) later.

Luo W., Conter L., Elsner R.A., Smita S., Weisel F., Callahan D., Wu S., Chikina M, & Shlomchik M. (2023). IL-21R signal reprogramming cooperates with CD40 and BCR signals to select and differentiate germinal center B cells. Science immunology, 8(80), eadd1823.

+ Open protocol

+ Expand

Peptide-based Immunotherapeutic Strategies

Check if the same lab product or an alternative is used in the 5 most similar protocols

The E7^44–62 peptide, Q19D (QAEPDRAHYNIVTFCCKCD); E7^49–57 peptide, R9F (RAHVYNIVTIF); E6^43–57 peptide, Q15L (QLLRREVYDFAFRDL); and E6^49–58 peptide, V10C (VYDFAFRDLC) were purchased from Elim Biopharma. The glycolipid α-galactosylceramide (αGalCer) adjuvant was purchased from DiagnoCine. APC-labeled H-2D^b epitope E7^49–57 (RAHYNIVTF)-containing tetramer was procured from the MHC tetramer production facility at Baylor College of Medicine (Houston, TX). The tumor-infiltrating lymphocytes (TIL) were analyzed by multi-parametric flow cytometry using the antibodies described in the Supplementary Methods. The following antibodies for immunotherapy were purchased from BioXcell and used at the concentrations shown: α4–1BB (LOB12.3 at 350 μg per dose), αCD40 (FGK4.5 at 100 μg per dose), αCTLA-4 (9H10 at 100 μg per dose), αPD-1 (RMP1–14 at 250 μg per dose), and αOX-40 (OX-86 at 100 μg per dose).

Dorta-Estremera S., Chin R.L., Sierra G., Nicholas C., Yanamandra A.V., Nookala S.M., Yang G., Singh S., Curran M.A, & Sastry K.J. (2018). Mucosal HPV E6/E7 Peptide Vaccination in Combination with Immune Checkpoint Modulation Induces Regression of HPV+ Oral Cancers. Cancer research, 78(18), 5327-5339.

+ Open protocol

+ Expand

In vitro and in vivo DC stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

For in vitro DC stimulation, 500,000 Flt3L-cultured cells were plated in complete IMDM and stimulated with αCD40 (10 μg ml⁻¹, FGK4.5, BioXCell), poly(I:C) (4 μg ml⁻¹, Sigma-Aldrich) or combined αCD40 + poly(I:C). Cells were stimulated for 24 h and then prepared for surface staining and flow cytometry. For in vivo DC stimulation, mice were injected i.p. with αCD40 (200 μg, BioXCell), poly(I:C) (150 μg, Sigma-Aldrich) or combined αCD40 + poly(I:C) in 150 αl PBS. After 24 h, spleens and SDLNs were harvested from mice and prepared for DC analysis as described above.

Wu R., Ohara R.A., Jo S., Liu T.T., Ferris S.T., Ou F., Kim S., Theisen D.J., Anderson DA I.I.I., Wong B.W., Gershon T., Schreiber R.D., Murphy T.L, & Murphy K.M. (2022). Mechanisms of CD40-dependent cDC1 licensing beyond costimulation. Nature immunology, 23(11), 1536-1550.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!