Adjacent liver and tumor samples were collected from SB, KPPC, and FAC tumor-bearing C57BL/6J mice. The samples were minced and dissociated into single cells using the Miltenyi mouse tumor dissociation kit (130-096-730) for tumor samples or the mouse liver dissociation kit (130-105-807) for adjacent uninvolved liver samples. Cell digestion of both tumor and adjacent normal samples was strained through a 70-μm cell strainer, and blood cells were eliminated using a red blood cell lysis buffer. Single cells were stained with viability dye (564406; BD Horizon) and PE/cyanine7 anti-mouse CD45 antibody (103113; BioLegend). CD45-positive viable cells were sorted using the BD FACSMelody Cell Sorter. Cells were stained with TotalSeq-C Mouse Universal Cocktail (V1.0, 199903; BioLegend). Finally, the stained single-cell suspension was processed and sequenced at the Genome Analysis Core (Mayo Clinic).
Pe cyanine7 anti mouse cd45 antibody
Manufactured by BioLegend
Sourced in United States
The PE/Cyanine7 anti-mouse CD45 antibody is a fluorescently-labeled antibody that specifically binds to the CD45 surface antigen on mouse cells. It can be used to identify and distinguish different types of hematopoietic cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria 3 sorter, by BD (3 mentions)
Apc anti mouse tnf α antibody, by BioLegend (3 mentions)
Brilliant violet 421 anti mouse human cd11b antibody, by BioLegend (3 mentions)
Fitc anti mouse il 6 monoclonal antibody mp5 20f3, by Thermo Fisher Scientific (2 mentions)
Fitc anti human ace2 antibody nbp2 7211f, by Novus Biologicals (2 mentions)
Alexa 647 anti iba1 antibody, by Cell Signaling Technology (2 mentions)
Annexin 5 fitc apoptosis detection kit bms500fi 20, by Thermo Fisher Scientific (2 mentions)
Mouse tumor dissociation kit, by Miltenyi Biotec (1 mentions)
Mouse liver dissociation kit, by Miltenyi Biotec (1 mentions)
Facsmelody cell sorter, by BD (1 mentions)
4 protocols using pe cyanine7 anti mouse cd45 antibody
1
Dissociation and Sorting of Liver and Tumor Cells
2
Multiparameter Immune Profiling of ZIKV-Infected Mice
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Isolated brain mononuclear cells from the mock- or ZIKV-infected mice in cell staining buffer (PBS with 1% FBS and 0.09% NaN3) were stained for 30 min with fluorescence-conjugated antibodies, namely, Brilliant Violet 421 anti-mouse/human CD11b antibody (101236; BioLegend, San Diego, CA), PE/Cyanine7 anti-mouse CD45 Antibody (103114; BioLegend), APC anti-mouse TNF-α antibody (506307; BioLegend), FITC anti-mouse IL-6 monoclonal antibody (MP5-20F3; 11-7061-82; eBioscience, San Diego, CA), PE anti-mouse IL-1β antibody (12-7114-82, eBioscience), and Alexa Fluor 488 anti-flavivirus group antigen antibody (4G2; NBP2-52709AF488; Novus Biologicals, Englewood, CO). The cells were then analyzed using a FACSAria III sorter (BD Biosciences, San Jose, CA), and data were analyzed using FlowJo software (BD Biosciences). All the fluorochromes were compensated for.
3
Flow Cytometry Analysis of Microglia
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Isolated brain mononuclear cells from the mock or SARS-CoV-2-infected mice in cell staining buffer (phosphate-buffered saline [PBS] with 1% FBS and 0.09% NaN3) were stained for 30 min with fluorescence-conjugated antibodies, namely, brilliant violet 421 anti-mouse/human CD11b antibody (101236, BioLegend, San Diego, CA, USA), PE/Cyanine7 anti-mouse CD45 antibody (103114, BioLegend), APC anti-mouse TNF-α antibody (506307, BioLegend), FITC anti-mouse IL-6 monoclonal antibody (MP5-20F3) (11-7061-82, eBioscience, San Diego, CA, USA), FITC anti-human ACE2 antibody (NBP2-7211F, Novus Biologicals, Centennial, CO, USA), and Alexa 647 anti-Iba1 antibody (78060S, Cell Signaling Technology, Danvers, MA, USA). Cells were then analyzed by FACSAria III sorter (BD Biosciences, San Jose, CA, USA), and data were analyzed by FlowJo software (BD Biosciences). All fluorochromes were compensated. The total leukocyte population was gated for microglia (CD11b+, CD45low). For annexin V staining, mock or SARS-CoV-2-infected HMC3 were stained with FITC-recombinant human annexin V, following the protocols of Annexin V-FITC Apoptosis Detection kit (BMS500FI-20, eBioscience).
4
Flow Cytometry Analysis of Microglia
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Isolated brain mononuclear cells from the mock or SARS-CoV-2-infected mice in cell staining buffer (phosphate-buffered saline [PBS] with 1% FBS and 0.09% NaN3) were stained for 30 min with fluorescence-conjugated antibodies, namely, brilliant violet 421 anti-mouse/human CD11b antibody (101236, BioLegend, San Diego, CA, USA), PE/Cyanine7 anti-mouse CD45 antibody (103114, BioLegend), APC anti-mouse TNF-α antibody (506307, BioLegend), FITC anti-mouse IL-6 monoclonal antibody (MP5-20F3) (11-7061-82, eBioscience, San Diego, CA, USA), FITC anti-human ACE2 antibody (NBP2-7211F, Novus Biologicals, Centennial, CO, USA), and Alexa 647 anti-Iba1 antibody (78060S, Cell Signaling Technology, Danvers, MA, USA). Cells were then analyzed by FACSAria III sorter (BD Biosciences, San Jose, CA, USA), and data were analyzed by FlowJo software (BD Biosciences). All fluorochromes were compensated. The total leukocyte population was gated for microglia (CD11b+, CD45low). For annexin V staining, mock or SARS-CoV-2-infected HMC3 were stained with FITC-recombinant human annexin V, following the protocols of Annexin V-FITC Apoptosis Detection kit (BMS500FI-20, eBioscience).
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