NK cells were treated with 20mM of PCI-34051 for 1 hour prior to cytokine stimulation. Cells were washed and resuspended in RPMI with 20% FBS, followed by co-culture with 5000 K562 cells or Jeko-1 cells at 1:10 effector to target ratios and cytolytic function assessed at 4 hour post co-culture. For antibody dependent cellular cytotoxicity (ADCC) experiments, Jeko-1 cells were pretreated with 10μg of Rituximab for 1 hour (#A1049, BioVision), washed twice to remove excess Rituximab, and co-cultured with stimulated NK cells for 4 hours. Cr-51 labeling and release was measured as previously described [16 (link)].
Rituximab
Manufactured by Abcam
Sourced in United States
Rituximab is a monoclonal antibody that targets the CD20 antigen expressed on the surface of B cells. It is a widely used laboratory reagent in immunology research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Cellquest software, by BD (1 mentions)
Anti his tag, by Abcam (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Infliximab, by Merck Group (1 mentions)
Maraviroc, by Merck Group (1 mentions)
Ici 118 551 hydrochloride, by Merck Group (1 mentions)
Staphylococcal enterotoxin a sea, by Merck Group (1 mentions)
Tram 34, by R&D Systems (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
3 protocols using rituximab
1
NK Cell-Mediated Cytotoxicity Assay
2
Quantifying scFv Binding to CD20 on Raji Cells
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In this experiment, the ability of selected scFv to bind to the native CD20 on Raji cells was measured. The Rituximab, a commercial anti-CD20 monoclonal antibody, was used as a positive control. First, cells were detached using 0.25% Trypsin–EDTA and counted using Hemocytometer. Then, for each antibody staining, 1 × 106 cells were washed twice with 1% PBS-BSA. All the antibody staining were performed for 1 h at RT in darkness. Cells were washed twice after each antibody staining. Next, cells were stained with 100 nM either Rituximab (Kindly provided by Tehran University Hospital) or scFv. Afterward, the secondary antibody was added to each tube, anti-Human (Abcam, USA) and anti-his tag (Abcam, USA) FITC conjugated for Rituximab and scFv stained samples, respectively. Next, one sample of each cell line was stained with Isotype as a negative control. Finally, the cells were analyzed by FACS-Calibur flow cytometer and CellQuest software (Becton Dickinson, San Jose, CA, USA).
3
Characterization of Immune Cell Responses
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Calcein-AM (#C3100) was purchased from ThermoFisher Scientific, staphylococcal enterotoxin A (SEA, #S9399), Infliximab (#Y0002047), ICI 118,551 hydrochloride (#127-5MG) and Maraviroc (#PZ0002) were from Sigma Aldrich. TRAM 34 (#2946) was purchased from R&D Systems and Venetoclax (ABT-199, #ab217298) from Abcam, Rituximab (lot oN7370B16) was from the local pharmacy (Universitätsklinikum Homburg, Hoffmann La Roche). All other chemicals and reagents not specifically mentioned were from Sigma Aldrich, ThermoFisher Scientific.
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