The MojoSort Mouse NK Cell Isolation Enrichment Kit (BioLegend, 480050) was used to isolate untouched NK cells. Adoptive NK cell transfer into NSG hosts was performed via tail vein injection with a 1:1 donor:recipient ratio. Magnetic enrichment was validated by FC prior to ACT for each purification. Typical purifications were 85–95% pure. Flow cytometry (FC) was performed by treating cells with an Fc-Shield antibody cocktail (αCD16/αCD32) in FACS buffer (Hank’s balanced salt solution [HBSS], 5 mM EDTA, and 2% fetal calf serum [FCS]) using the following antibodies (1:400 dilution): anti-mouse CD45− PE-Cy7 (BD Biosciences, 559864), NK 1.1-BV 785 (BD Biosciences, 108749), and NKP46- BV 421 (BD Biosciences, 562850) for 60 min on ice. Cells were washed and centrifuged at 400 × g for 7 min twice. Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) and analyzed on a FACSymphony A5 cytometer (BD Biosciences, San Jose, CA, USA). Data were analyzed with FlowJo version 10 (Tree Star, Ashland, OR, USA).
Nkp46 bv 421
Manufactured by BD
The NKP46-BV 421 is a laboratory equipment product. It is a fluorescently-labeled antibody that binds to the NKP46 receptor, which is expressed on the surface of natural killer (NK) cells. The BV 421 dye is used to label the antibody, enabling detection and analysis of NKP46-positive cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse cd45 pe cy7, by BD (1 mentions)
Facsymphony a5 cytometer, by BD (1 mentions)
Cd69 fitc, by BD (1 mentions)
Cd69 apc, by BD (1 mentions)
Ly6c apc, by BioLegend (1 mentions)
Cd4 apc cy7, by BD (1 mentions)
Pd 1 pe, by BD (1 mentions)
Cd8 pe cy5, by BD (1 mentions)
Ly6c apc, by BD (1 mentions)
Cd8a pe cy7, by BD (1 mentions)
Mhc 2 bv650, by BD (1 mentions)
Nkg2d pe cy7, by BD (1 mentions)
Foxp3 fitc, by BD (1 mentions)
Pd l1 bv421, by BD (1 mentions)
F4 80 pe cy7, by BD (1 mentions)
Collagenase 1, by Merck Group (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Facscanto 2, by BD (1 mentions)
Cd8 pe cy7, by BD (1 mentions)
Cd44 fitc, by BD (1 mentions)
2 protocols using nkp46 bv 421
1
Isolation and Adoptive Transfer of Murine NK Cells
2
Profiling Circulating Immune Cells Post-Cardiac Bleed
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Blood obtained from cardiac bleeds was used to profile circulating immune cells after red blood cell lysis (155 mM NH4Cl, 10 mM KHCO3, 0.1 mM EDTA, pH 7.3). Cells were stained with the following antibodies: CD8a-PE-Cy7 (53-6.7), CD4-APC-Cy7 (GK1.5), CD69-APC (H1.2F3), CD279-PE (J43), Ly6G-BV421 (1A8), B220 Pe-Cy5, CD103 bv510, CD11b BV605, CD11c Percpm, CD206 APC, CD25 APC, CD27 A-Cy7, CD4 A-Cy7, CD4 Pe-Cy7, CD44 FITC, CD62L BV450, CD69 FITC, CD69 Pe-Cy7, CD8 A-Cy7, CD8 APC, CD8 Pe-Cy5, CD8 Pe-Cy7, CD80 PE, F4/80 Pe-Cy7, FOXP3 FITC, H2-Kd FITC, IFNAR1 PE, IFNg PE, Ly6C-APC, Ly6G BV711, MHC-II BV650, NKG2D Pe-Cy7, NKp46 BV421, PD1 PE, PDL1 BV421, Rat IgG2a FITC, TCRbeta BV510, TNFa FITC (all from BD Biosciences) and Ly6C-APC (HK1.4) (Biolegend). Primary tumors were mechanically and enzymatically digested with 1 mg/mL collagenase I (Sigma) and 30 μg/mL DNAse I (Sigma) at 37 °C to obtain a single cell suspension before red blood cell lysis. Analysis of tumor-infiltrating lymphocytes was done as above. Analysis of immune cell populations was performed by flow cytometry using a FACSCanto II (BD Biosciences, USA). Data was analyzed using Flowjo software (TreeStar, USA).
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