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Anti mouse cd28

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Anti-mouse CD28 is a laboratory reagent used for the detection and analysis of the CD28 cell surface receptor in mouse samples. It is a monoclonal antibody that binds specifically to the CD28 protein, which is expressed on the surface of T cells and plays a crucial role in T cell activation and costimulation.

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Lab products found in correlation

Anti mouse cd3, by BioXCell (2 mentions) Interleukin 2 (il 2), by R&D Systems (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) 2 nbdg, by Thermo Fisher Scientific (1 mentions) Anti human cd3, by BioLegend (1 mentions) Anti human cd28, by BioLegend (1 mentions) Cfse proliferation dye, by Thermo Fisher Scientific (1 mentions) Aqua live dead viability dye, by Thermo Fisher Scientific (1 mentions) Cfse cell division tracker kit, by BioLegend (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Mojosort mouse cd8 t cell isolation kit, by BioLegend (1 mentions) Live dead fixable violet dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Percp cyanine5.5 conjugated anti mouse cd8a antibody, by BioLegend (1 mentions) Cytoflex lx, by Beckman Coulter (1 mentions) Fetal bovine serum (fbs), by Sartorius (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)

4 protocols using anti mouse cd28

1

T cell proliferation assay with macrophages

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The CD4+ and CD8+ T cells were magnetically enriched (Biolegend, USA) from the spleen of 6-week-old C57BL/6 mice. Separated T cells were seeded into 96-well plate pre-coated with 2 μg/mL anti-mouse CD3 (BioXcell, USA) in IMDM (Hyclone, USA) containing 2 μg/mL anti-mouse CD28 (BioXcell, USA), 10% fetal bovine serum, 100 U/mL interleukin-2 (R&D system, USA) and 2 mM l-glutamine (Gibco, USA). After being labeled with 1 μM CFSE (Thermo, USA), 3 × 106 T cells were co-cultured with peritoneal macrophages with indicated treatment at a ratio of 2:1. After incubation for 3 days, cells were collected, and FACS analyzed the CFSE signal.
Sun J.L., Zhang N.P., Xu R.C., Zhang G.C., Liu Z.Y., Abuduwaili W., Wang F., Yu X.N., Shi X., Song G.Q., Wu H., Liu T.T., Shen X.Z., Deng B., Weng S.Q., Dong L, & Zhu J.M. (2021). Tumor cell-imposed iron restriction drives immunosuppressive polarization of tumor-associated macrophages. Journal of Translational Medicine, 19, 347.
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2

Cytokine-Mediated T Cell Polarization

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Mouse and human cytokines used for in vitro polarizations as well as anti-human CD3 and anti-human CD28 were purchased from Biolegend. Anti-mouse CD3, anti-mouse CD28 and mouse and human neutralizing antibodies to IL-4 and IFN-γ were purchased from BioXCell. Anti-CK2α and anti-CK2β antibodies for detection by flow cytometry were purchased from AbCam and Calbiochem, respectively. Secondary antibodies for flow cytometry were purchased from Jackson ImmunoResearch. Flow cytometry antibodies against mouse CD4, IL-17A, IFN-γ, CD25 and CD45.1 and human CD3, CD4 and Foxp3 were purchased from Biolegend. Flow cytometry antibodies against mouse CD44, Foxp3 and GM-CSF and human IL-17A were purchased from eBioscience. Aqua Live/Dead Viability dye, CFSE proliferation dye and 2-NBDG were purchased from ThermoFisher Scientific. Flow cytometry antibodies and isotype controls for phosphorylated S6, Akt, STAT3, STAT5 and SMAD2/3 were purchased from Cell Signaling. Immunoblotting antibodies against phosphorylated T389, S371 and total S6 Kinase p70 and phosphorylated S129, S473 and total Akt were purchased from Cell Signaling, and antibody against mouse β-actin was purchased from abcam.
Gibson S.A., Yang W., Yan Z., Liu Y., Rowse A.L., Weinmann A.S., Qin H, & Benveniste E.N. (2017). Protein Kinase CK2 Controls the Fate Between Th17 Cell and Regulatory T Cell Differentiation CK2 Regulates the Th17/Treg Axis. Journal of immunology (Baltimore, Md. : 1950), 198(11), 4244-4254.
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3

Antibody Reagents for Murine NKG2A and Ly-49

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Anti-mouse NKG2A (16A11,
purified) was purchased from BioLegend. Anti-mouse NKG2A (20D5, purified)
and anti-mouse CD28 (37.51, purified) were purchased from BioXcell.
Anti-mouse Ly-49C and Ly-49I (5E6, purified) were purchased from BD
Bioscience.
Ren J., Jo Y., Picton L.K., Su L.L., Raulet D.H, & Garcia K.C. (2022). Induced CD45 Proximity Potentiates Natural Killer Cell Receptor Antagonism. ACS Synthetic Biology, 11(10), 3426-3439.
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4

Isolation and Activation of Murine CD8+ T Cells

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A round-bottom 96-well plate was first prepared by incubation with 100 μl PBS supplemented with 1 μg/mL anti-mouse CD3ε (BioXCell, BE0001-1-5MG) and anti-mouse CD28 (BioXCell, BE0015-1-5MG) the day before CD8+ T-cell isolation. The supernatant was discarded before use.
CD8+ T cells were isolated from the spleen tissue of adult male C57BL/6 mice with a MojoSort Mouse CD8 T Cell Isolation Kit (BioLegend, 480035) according to the standard protocol. The isolated CD8+ T cells were first incubated with reagents from a CFSE Cell Division Tracker Kit (BioLegend, 423801) according to the standard protocol and then resuspended in RPMI 1640 medium (Gibco, 11875093) supplemented with 10% FBS (Biological Industries, 04-001-1A) and 1% Pen Strep (Gibco, 15140122). Then, 20 ng/mL mouse IL2 (Novoprotein, CK24) and IL7 (Novoprotein, CC73) were added, and the concentration of cells was 106/mL.
The isolated CD8+ T cells were then incubated in the precoated 96-well plates for 72 hours. After that, the cells were collected and stained with a LIVE/DEAD Fixable Violet Dead Cell Stain Kit (Invitrogen, L34963), PerCP/Cyanine 5.5-conjugated anti-mouse CD8a antibody (BioLegend, 100733) and PE-conjugated anti-mouse IFN-γ antibody (BioLegend, 163503) as described above. The prepared cell suspensions were analyzed on a CytoFLEX LX from Beckman Coulter.
Wang R., Zhang Y., Guo S., Pei S., Guo W., Wu Z., Wang H., Wang M., Li Y., Zhu Y., Meng L.H., Lang J., Jin G., Xiao Y., Hu L, & Kong X. (2023). Single-cell RNA sequencing reveals the suppressive effect of PPP1R15A inhibitor Sephin1 in antitumor immunity. iScience, 26(2), 105954.
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