Soluble anti cd28 clone 37

Manufactured by BioXCell

Sourced in United States

Soluble anti-CD28 (clone 37.51) is a laboratory reagent used for cell biology research. It is an antibody that binds to the CD28 receptor on the surface of T cells. This interaction can provide a costimulatory signal that enhances T cell activation and proliferation.

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Lab products found in correlation

Anti cd3 clone 145 2c11, by BioXCell (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) Human il 2, by Thermo Fisher Scientific (1 mentions) Ot 2 tcr transgenic mice, by Jackson ImmunoResearch (1 mentions) Anti cd3 clone 2c11, by BioXCell (1 mentions) Anti cd3, by BioXCell (1 mentions) Il 17, by BD (1 mentions) Mouse na ve cd4 t cells isolation kit, by Miltenyi Biotec (1 mentions) Anti ifn γ, by BioXCell (1 mentions) Anti il 4, by BioXCell (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Il 12, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by BioXCell (1 mentions) Soluble anti cd28, by BioXCell (1 mentions) Golgiplug, by BD (1 mentions)

Close spelling variants of this product

Soluble anti-mouse CD28 (clone 37.51, Anti-CD28 (clone 37.51, Soluble anti-CD28 Anti-CD28 (37.51, Clone 37.51 Anti-CD28

3 protocols using soluble anti cd28 clone 37

Isolation and Activation of Mouse CD4+ T Cells

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CD4⁺ T cells were isolated from mouse spleens using a negative selection kit according to the manufacturer’s protocol (either #130-104-454 from Miltenyi Biotech (Bergisch Gladbach, Germany) or #19852 from Stem Cell Technologies (Vancouver, Canada)). Purity of isolated cells was typically >95%, and cells were resuspended in complete RPMI medium (10% FCS, 2mM glutamine (#25030–149, Gibco), 50U/ml Penicillin + 50μg/ml streptomycin (#15070–063, Gibco) and 2-betamercaptethanol (21985–023, Gibco)). Cells were activated in plates pre-coated with 2ug/ml of anti-CD3 (clone 145–2C11, BioXcell, West Lebanon, NH) at 2×10⁵ per well of a 96 well plate and 5×10⁵ per well of a 48 well plate in the presence of 1ug/ml soluble anti-CD28 (clone 37.51, BioXcell, West Lebanon, NH). After 3 days, the cells were harvest, split into half and seeded into a new plate with 25U/ml Human IL-2 (Peprotech). Cells were harvested and assayed at the indicated timepoints.

West E.E., Merle N.S., Kamiński M.M., Palacios G., Kumar D., Wang L., Bibby J.A., Overdahl K., Jarmusch A.K., Freeley S., Lee D.Y., Thompson J.W., Yu Z.X., Taylor N., Sitbon M., Green D.R., Bohrer A., Mayer-Barber K.D., Afzali B., Kazemian M., Scholl-Buergi S., Karall D., Huemer M, & Kemper C. (2023). Loss of CD4+ T cell-intrinsic arginase 1 accelerates Th1 response kinetics and reduces lung pathology during influenza infection. Immunity, 56(9), 2036-2053.e12.

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In Vitro Activation of CD4+ T Cells

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All in vitro experiments were conducted in accordance with UCLA’s institutional policy on humane and ethical treatment of animals following protocols approved by the Animal Research Committee. Five- to eight-week-old OT-II TCR transgenic mice (Jackson Labs) were used for all experiments. Cell-culture media was RPMI supplemented with 10% heat-inactivated FBS, 1% penicillin/streptomycin, 1% sodium pyruvate, 1% HEPES buffer, 0.1% μM 2-mercaptoethanol. CD4+ T cells were purified using negative selection enrichment kits (Stem Cell Technologies).
In vitro activation of CD4+ T cells was performed by culturing 1×10⁶ cells/mL in tissue culture-treated 24-well plates that were pre-coated with anti-CD3 (clone 2C11; Bio X Cell) at a concentration of 10 μg/mL plus addition of 2 μg/mL soluble anti-CD28 (clone 37.51; Bio X Cell).

Majedi F.S., Hasani-Sadrabadi M.M., Thauland T.J., Li S., Bouchard L.S, & Butte M.J. (2020). T-cell activation is modulated by the 3D mechanical microenvironment. Biomaterials, 252, 120058.

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Differentiation of Naïve CD4+ T cells

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Naïve CD4⁺ T cells were purified from the splenocytes of 6-7-week-old EGR2^-/-B6/lpr and control mice by using a mouse naïve CD4⁺ T cells isolation kit from Miltenyi Biotec as we previously reported (16 (link)). For Th1 differentiation, naïve CD4⁺ T cells (1.5x10⁶) were plated in 48-well plate and cultured with 2 μg/ml plate-bound anti-CD3 (clone 145-2C11, Bio X cell, Lebanon, NH, USA), 1 μg/ml soluble anti-CD28 (clone 37.51, Bio X cell), 5ng/ml IL-2 (eBioscience), 10ng/ml IL-12 (eBioscience) and 10 μg/ml anti-IL4 (Bio X cell) for 3 days. For Th17 differentiation, naïve CD4⁺ T cells (1.5x10⁶) were plated in 48-well plate and cultured with 2 μg/ml plate-bound anti-CD3, 1 μg/ml soluble anti-CD28, 20ng/ml IL-6, 3ng/ml hTGFβ1, 10 μg/ml anti-IFNγ (Bio X cell) and 10 μg/ml anti-IL4 for 5 days. The differentiated cells were stimulated with PMA (50 ng/ml), ionomycin (1 μg/ml) plus protein transporter inhibitor (BD GolgiPlug) for 5 hours, and then collected for intracellular flow cytometry analysis of IFNγ- and IL-17- expressing cells.

Dai R., Wang Z., Heid B., Eden K., Reilly C.M, & Ahmed S.A. (2022). EGR2 Deletion Suppresses Anti-DsDNA Autoantibody and IL-17 Production in Autoimmune-Prone B6/lpr Mice: A Differential Immune Regulatory Role of EGR2 in B6/lpr Versus Normal B6 Mice. Frontiers in Immunology, 13, 917866.

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