human cd4 cell recovery column kit  (Cedarlane)

Journal: Cell reports

Article Title: Calcineurin is an adaptor required for assembly of the TCR signaling complex

doi: 10.1016/j.celrep.2024.114568

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Article Snippet: Human CD4 + T cells were isolated from buffy coats of healthy volunteers (NIH Blood Bank) using the Human CD4 cell recovery column kit (Cedarlane), according to the manufacturer’s instructions.

Techniques: Purification, Recombinant, Protease Inhibitor, Cell Recovery, In Situ, Plasmid Preparation, Control, Software, Luminex

Journal: Vaccines

Article Title: Disc Large Homolog 1 Is Critical for Early T Cell Receptor Micro Cluster Formation and Activation in Human T Cells

doi: 10.3390/vaccines9121446

Figure Lengend Snippet: DLG1 knockdown inhibits TCR-MC formation and subsequent proximal signaling. ( A , B ). Microcluster analysis. Scale bars, 10 μm. ( A ) Jurkat cell analysis. Microclusters of ZAP-70–YFP (green) and phosphorylated TCR-ζ (red) were visualized in immunostained Jurkat cells stably expressing ZAP-70–YFP, pretreated with control or DLG1-specific siRNA. Cells were dropped onto coverslips coated with anti-CD3 (stimulated) or anti-CD45 (unstimulated) and visualized after 5 min. Quantitative data for specific signals’ intensities (using Image J; collected from eight fields) are graphically represented. ( B ) Primary human CD4 + T cells. Microclusters of phosphorylated Y493 ZAP-70 (green) and phosphorylated TCR-ζ (red) were visualized in immunostained human T cells, pretreated with control or DLG1-specific siRNA. Cells were dropped onto coverslips coated with anti-CD3+anti-CD28 (stimulated) or anti-CD45 (unstimulated) and visualized after 5 min. Quantitative data for specific signals’ intensities (using Image J; collected from eight fields) are graphically represented. ( C , D ). Immunoblot analysis of lysates from Jurkat cells ( C ) and human T cells ( D ) pretreated with control or DLG1-specific siRNA and then activated with soluble anti-CD3 (monoclonal antibody OKT3) and antibody to mouse immunoglobulin G (IgG), followed by probing for phosphorylated and total proteins. Data are representative of three experiments. Student’s unpaired t tests were used for ( A , B ). ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CD4 recovery column kits were from Cedarlane Laboratories Limited, Burlington, ON, Canada.

Techniques: Knockdown, Cell Analysis, Stable Transfection, Expressing, Control, Western Blot