Granzyme Monoclonal Antibody (NGZB), PE-Cyanine7, eBioscience™

Protocol tips

Upstream tips	Protocol tips	Downstream tips
For flow cytometry analysis, BD GolgiPlug Protein Transport Inhibitor (BD Biosciences, San Jose, CA) was added, and cells were cultured for 10 h in 5% CO2 at 37 °C.	After incubation, cells were washed with FACS buffer (PBS with 2% FBS) and stained with Acqua Live/Dead (Invitrogen, USA) for 10 min at 4 °C for dead cell exclusion. Cells were washed and stained for surface molecules for 30 min at 4 °C, fixed and permeabilized according to the manufacturer’s instructions (Cytofix/Cytoperm, BD Biosciences, USA). After washing, cells were incubated with antibodies against intracellular antigens for 30 min at 4 °C. Cells were then washed and suspended in 200 μL of FACS buffer for cytometry analysis.	Data were collected using an LSR II (BD Immunocytometry Systems, USA) with Diva (BD Biosciences, USA). At least 100,000 gated events were acquired.

Upstream tips
For flow cytometry analysis, BD GolgiPlug Protein Transport Inhibitor (BD Biosciences, San Jose, CA) was added, and cells were cultured for 10 h in 5% CO2 at 37 °C.
Protocol tips
After incubation, cells were washed with FACS buffer (PBS with 2% FBS) and stained with Acqua Live/Dead (Invitrogen, USA) for 10 min at 4 °C for dead cell exclusion. Cells were washed and stained for surface molecules for 30 min at 4 °C, fixed and permeabilized according to the manufacturer’s instructions (Cytofix/Cytoperm, BD Biosciences, USA). After washing, cells were incubated with antibodies against intracellular antigens for 30 min at 4 °C. Cells were then washed and suspended in 200 μL of FACS buffer for cytometry analysis.
Downstream tips
Data were collected using an LSR II (BD Immunocytometry Systems, USA) with Diva (BD Biosciences, USA). At least 100,000 gated events were acquired.

Publication protocol

For flow cytometry analysis, BD GolgiPlug Protein Transport Inhibitor (BD Biosciences, San Jose, CA) was added, and cells were cultured for 10 h in 5% CO2 at 37 °C. After incubation, cells were washed with FACS buffer (PBS with 2% FBS) and stained with Acqua Live/Dead (Invitrogen, USA) for 10 min at 4 °C for dead cell exclusion. Cells were washed and stained for surface molecules for 30 min at 4 °C, fixed and permeabilized according to the manufacturer’s instructions (Cytofix/Cytoperm, BD Biosciences, USA). After washing, cells were incubated with antibodies against intracellular antigens for 30 min at 4 °C. Cells were then washed and suspended in 200 μL of FACS buffer for cytometry analysis. Data were collected using an LSR II (BD Immunocytometry Systems, USA) with Diva (BD Biosciences, USA). At least 100,000 gated events were acquired. Representative FACS density plots showing the gate strategy for the identification of IL-1β within CD11c+MHCIIhigh and iNOS within F4/80+ gated on live CD45+ leucocytes in the trigeminal ganglia and spleen from a single HSV1-infected wild type mouse were done on Additional file 1: Figure S1 and Additional file 2: Figure S2, respectively.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Flow cytometry Anti-bodies Mouse - Granzyme B using Granzyme Monoclonal Antibody (NGZB), PE-Cyanine7, eBioscience™ from eBioscience.

Manufacturer protocol

Download the product protocol from eBioscience for Granzyme Monoclonal Antibody (NGZB), PE-Cyanine7, eBioscience™ below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Flow cytometry Anti-bodies Mouse - Granzyme B using Granzyme Monoclonal Antibody (NGZB), PE-Cyanine7, eBioscience™ from eBioscience. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.