Leukocyte-enriched peripheral blood samples of human healthy donors were obtained from the American Red Cross. Primary NK cells were isolated using the MACSxpress® NK Cell Isolation Kit (Miltenyi Biotec) and Erythrocyte Depletion Kit (Miltenyi Biotec). Enriched NK cells were approximately 99% pure, which was confirmed by flow cytometry using anti-CD3 and anti-CD56 antibodies (BD Biosciences). Primary human NK cells were used for the experiments in this study unless otherwise indicated that the NK-92 cell line was used for specific experiments. The protocols for human specimen collection were approved by the IRBs of The Ohio State University.
Anti cd3 and anti cd56 antibodies
Manufactured by BD
Sourced in United States
Anti-CD3 and anti-CD56 antibodies are laboratory reagents used for the identification and enumeration of specific cell types in biological samples. Anti-CD3 antibodies recognize the CD3 antigen, a complex of membrane-bound proteins associated with the T-cell receptor, and are used to identify T lymphocytes. Anti-CD56 antibodies recognize the CD56 antigen, also known as neural cell adhesion molecule (NCAM), and are used to identify natural killer (NK) cells. These antibodies are commonly used in flow cytometry and other immunoassay applications.
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3 protocols using anti cd3 and anti cd56 antibodies
1
Isolation and Validation of Primary Human NK Cells
2
Isolation and Validation of Primary Human NK Cells
3
Characterization of NK Cells in Hepatitis B
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Peripheral blood samples and clinical assessments were obtained during routine follow-ups of hepatitis patients. CHB patients in the IT phase were admitted to the Qilu Hospital at Shandong University. All patients were negative for other viral infections, including hepatitis D virus, HCV and HIV, and had no autoimmune liver diseases. An age- and sex-matched control group was composed of healthy subjects. The clinical characteristics of these samples are described in Table 1 . In accordance with the Ethics Committee of Shandong University, informed consent was acquired from all participants.
Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll gradient separation after centrifugation. NK cells were purified by negative selection using a human NK cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions. Cell purity was determined by flow cytometry with anti-CD3 and anti-CD56 antibodies (BD Pharmingen, San Jose, CA, USA) and the purity of the NK cells (CD3−CD56+) was determined to be >95%.
Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll gradient separation after centrifugation. NK cells were purified by negative selection using a human NK cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions. Cell purity was determined by flow cytometry with anti-CD3 and anti-CD56 antibodies (BD Pharmingen, San Jose, CA, USA) and the purity of the NK cells (CD3−CD56+) was determined to be >95%.
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