PBMC was stained using Zombie Live/dead fixable cell stain (Thermo Fisher Scientific, Waltham, MA, USA) prior to staining with specific antibodies. Antibodies used were CD3 BUV496 (BD Bioscience), CD19 PeCy7 (BD Bioscience), CD56 BB700 (BD Bioscience), CD16 BV650 (BioLegend), CD96 BV421 (BD Bioscience), TIGIT BUV395 (BD Bioscience), DNAM1 (CD226) BUV805 (BD Bioscience). PBMC was stained in Live/Dead cell stain for 15 min in PBS at RT, washed in FACS buffer (PBS + 2% FCS), and blocked in Human Fc block (BD Bioscience) for five minutes at RT. Cell staining was performed in FACS buffer for 30 min at 4 °C, followed by two washes in FACS buffer, and fixation in 2% paraformaldehyde (PFA, Electron Microscopy Sciences). Samples were acquired on a LSR Fortessa (BD Bioscience) flow cytometer and analysed using FlowJo software (BD Bioscience). Flow cytometry gating is shown in Additional file 1 : Fig. S1. NK cells were defined as CD3-CD19-CD56 + viable single lymphocytes. NK cell subsets were defined as mature (CD16 + CD56mid), regulatory (CD16-CD56mid) and immature (CD16-CD56hi). Expression of DNAM1, TIGIT and CD96 was calculated using the geometric mean of the mature, regulatory and immature NK cell subsets.
Cd56 bb700
Manufactured by BD
The CD56 BB700 is a laboratory instrument used for the detection and analysis of CD56, a cell surface protein. It is designed to provide accurate and reliable data on the expression of CD56 in various biological samples. The CD56 BB700 utilizes advanced flow cytometry technology to enable researchers to quantify and characterize CD56-positive cells. This product is intended for research use only and its core function is to facilitate the analysis of CD56 expression.
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Lab products found in correlation
Human fc block, by BD (1 mentions)
Tigit buv395, by BD (1 mentions)
Cd3 buv496, by BD (1 mentions)
Cd16 bv650, by BioLegend (1 mentions)
Cd19 pe cy7, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Easysep human nk cell isolation kit, by STEMCELL (1 mentions)
Cd3 bv421, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
2 protocols using cd56 bb700
1
Multiparameter Flow Cytometry of NK Cells
2
Isolation and Purification of NK Cells
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PBMC from HC were stained for 30 min at 4°C with CD3BV421 and CD56BB700 (BD Biosciences) and subsequently were fluorescence-activated cell sorting (FACS)-sorted with FACS Aria III (BD Biosciences). LIVE/DEAD® Fixable Near-IR Dead Cell Stain Kit (Thermo Fisher Scientific) was used according to the manufacturer’s instructions to determine cell viability. Sorted NK cells exhibited a purity >98%. In a second experiment, NK cells were enriched by immunomagnetic negative selection from freshly PBMC using the EasySep™ Human NK Cell Isolation kit according to the manufacturer’s instructions (STEMCELL Technologies, Vancouver, Canada). In this case, NK cell purity was >95% (data not shown).
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