αcd28 37.51

Manufactured by BioXCell

αCD28 (37.51) is a laboratory reagent used in cell culture experiments. It functions as an antibody that binds to the CD28 receptor on T cells, which is an important costimulatory molecule involved in T cell activation and proliferation.

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Lab products found in correlation

Cd4 microbeads, by Miltenyi Biotec (2 mentions) αcd3 2c11, by BioXCell (2 mentions) Rapamycin, by Selleck Chemicals (1 mentions) Zstk474, by Merck Group (1 mentions) Recombinant il 2, by Thermo Fisher Scientific (1 mentions) Macs cell separation ms columns, by Miltenyi Biotec (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Interleukin 7 (il 7), by Thermo Fisher Scientific (1 mentions) Jes6 1a12, by BioXCell (1 mentions) Cd4 dynabeads, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions)

3 protocols using αcd28 37.51

Modulating T Cell Activation Pathways

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CD4⁺ T cells isolated from spleen and LN using CD4 microbeads
(Miltenyi). 1×10⁶ T cells cultured with platebound
α-CD3 (2C11) and α-CD28 (37.51) from BioXcell at 1μg/mL
each for 48 or 66 hrs. Inhibitors purchased and used as follows: ZSTK474
(1μM, Sigma), Rapamycin (10nM, Selleckchem), NFAT inhibitor (10μM,
Tocoris), Mek inhibitor PD0325901 (100nM, Peprotech) and Erk inhibitor FR180204
(10μM, Tocoris). In vitro T_R suppression
assays were performed as previous described (20 (link)). Chemotaxis assay performed as previously described (21 (link)).

Sullivan J.M., Höllbacher B, & Campbell D.J. (2018). Dynamic expression of Id3 defines the stepwise differentiation of tissue-resident regulatory T cells. Journal of immunology (Baltimore, Md. : 1950), 202(1), 31-36.

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T Cell Isolation and Activation

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VAT was pooled from two or three mice of the same genotype to ensure adequate cell number for culture. Cells were isolated as described above, and CD4⁺ T cells were enriched by incubating cells with CD4 MicroBeads and positively selecting with MACS cell separation MS columns (Miltenyi Biotec). Purified cells were resuspended in RPMI-C with 500 U/ml recombinant IL-2 (eBioscience). Cells were cultured for 48 h in 96-well flat-bottom plates with plate-bound αCD3 (2C11) and αCD28 (37.51) from Bio X Cell at 1 µg/µl, with or without the addition of plate-bound αICOS (C398.4A; BioLegend) at 2 µg/µl. Expression of CCR3 was assessed by flow cytometry as described above.

Mittelsteadt K.L., Hayes E.T, & Campbell D.J. (2021). ICOS signaling limits regulatory T cell accumulation and function in visceral adipose tissue. The Journal of Experimental Medicine, 218(6), e20201142.

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CFSE-Based T Cell Proliferation Assay

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Purified CD4⁺ T cells (CD4 T cells beads, Miltenyi or CD4 Dyna Beads, Invitrogen) were labeled with CFSE or CellTrace Volet (CTV) (Thermofisher) according to manufacturer’s protocol and cultured in RPMI 1640 complete medium supplemented with 10% fetal bovine serum (FBS, GIBCO Sigma), 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine (all from GIBCO) together with DC and irradiated feeder cells (40 Gy) in 2:1:1 ratio for 72 or 96h. Polyclonal CD4⁺ T cells were stimulated with αCD3 (2C11) and αCD28 (37.51) (BioXcell); OT-II cells with OVA_323-339 (ISQAVHAAHAEINEAGR) peptide (Pepscan) and Trp1 cells with Trp1_106-130 (SGHNCGTCRPGWRGAACNQKILTVR) peptide (Pepscan) at concentration indicated in the Figure 2. Cells were additionally supplemented with IL-2, IL-15 or IL-7 (Peprotech) at a concentration indicated in the Figure 2. Mouse CD4⁺ T cells were cultured with αCD25 (PC61, BioXcell) and αIL-2 (JES6-1A12, BioXcell), added to the culture 24h post stimulation with αCD3 and αCD28 in a concentration of 5 μg/ml.

Śledzińska A., Vila de Mucha M., Bergerhoff K., Hotblack A., Demane D.F., Ghorani E., Akarca A.U., Marzolini M.A., Solomon I., Vargas F.A., Pule M., Ono M., Seddon B., Kassiotis G., Ariyan C.E., Korn T., Marafioti T., Lord G.M., Stauss H., Jenner R.G., Peggs K.S, & Quezada S.A. (2020). Regulatory T Cells Restrain Interleukin-2- and Blimp-1-Dependent Acquisition of Cytotoxic Function by CD4+ T Cells. Immunity, 52(1), 151-166.e6.

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