CD4 + T cells and B cells were isolated from PBMCs using an isolation kit (Stem Cell Technologies) according to standard protocols. The selected cells were suspended in PBS solution containing 2% FBS and dropped onto polylysine-coated slides. The CD4 + T cells were stimulated by mouse anti-human CD3 (10 μg/ml; BioLegend) and mouse anti-human CD28 (10 μg/ml; BioLegend) at 37 °C. Stimulation was stopped by addition of PFA at 0, 5, 10, and 20 min. B cells were stimulated for different times with monobiotinylated goat-anti-human IgM + IgG Fab2 (Jackson Laboratories). Phalloidin-iFluor 488 (1:140) (Abcam) was used to stain F-actin, and DAPI (Beyotime) was used to stain nuclei. The mean fluorescence intensity of each protein was measured by the NIS-Elements AR 3.2 software.
Mouse anti human cd28
Manufactured by BioLegend
Mouse anti-human CD28 is a primary antibody that recognizes the CD28 surface antigen expressed on human T cells. CD28 is a co-stimulatory molecule that plays a crucial role in T cell activation and proliferation. This antibody can be used in flow cytometry and other immunoassays to detect and study CD28-positive cells.
Automatically generated - may contain errors
2 protocols using mouse anti human cd28
1
Visualizing T-cell and B-cell Activation
2
T Cell Activation by Thapsigargin and Anti-CD3/CD28
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T cells were activated using 2 μM thapsigargin (TG; MilliporeSigma) or with plate-bound mouse anti-human CD3 (10 μg/ml) and mouse anti-human CD28 (10 μg/ml) antibodies (BioLegend) as previously described (21 (link)). In experiments where IFNγ was measured using enzyme-linked immunosorbent assay (ELISA), some cells were activated with phytohemagglutinin (10 μg/ml). The activation protocol and activation times for each experiment are described in the individual figure legends.
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