Prior to perfusion, spleens were harvested from experimental mice. Single cell suspensions from spleens were treated with ACK erythrocyte lysis buffer. Mice were perfused with 25 mL of ice-cold PBS and brains and spinal cords were collected from perfused mice and dounce homogenized to obtain single cell suspensions. CNS cells were purified by centrifugation for 30 min in a 30% Percoll (GE Healthcare) solution as previously reported (25 (link)). Cells were incubated with the anti-Fc receptor antibody 2.4G2 prior to the addition of antibodies. The following antibodies were purchased from BD Biosciences: CD45-FITC, CD8α-APC-H7, CD19-APC-H7, CD19-BV510, B220-PE-TxRed, B220–PE-CF594, CD11b-AlexaFluor-700, MHC-v450. The following antibodies were purchased from eBioscience: MHCII-Pacific Blue, CD11c-PECy7. The following antibodies were purchased from BioLegend (San Diego, CA): CD138-PE, MHCII (I-A/I-E)-Pacific Blue, Thy1.1-PerCP, CD4-APC. Cells were acquired on a Gallios flow cytometer (Beckman Coulter) and analyzed with FloJo software (TreeStar) with doublets being excluded.
Mhc 2 pacific blue
Manufactured by Thermo Fisher Scientific
The MHC II-Pacific Blue is a fluorescently labeled antibody that specifically binds to the major histocompatibility complex class II (MHC II) molecule. It is used for the identification and characterization of cell populations expressing MHC II, which is commonly found on the surface of antigen-presenting cells such as dendritic cells, macrophages, and B cells.
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Lab products found in correlation
Cd45 fitc, by BD (1 mentions)
Cd11c pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd4 apc, by BioLegend (1 mentions)
Cd19 bv510, by BD (1 mentions)
Cd19 apc h7, by BD (1 mentions)
Percoll, by GE Healthcare (1 mentions)
Cd11b alexa fluor 700, by BD (1 mentions)
Cd8α apc h7, by BD (1 mentions)
Cd138 pe, by BioLegend (1 mentions)
Mhcii 1 a 1 e pacific blue, by BioLegend (1 mentions)
Thy1.1 percp, by BioLegend (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Flojo software, by Tree Star (1 mentions)
Signr1 apc, by Thermo Fisher Scientific (1 mentions)
Anti mouse cd11c pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd4 pe cy5, by BD (1 mentions)
Ly6c apc, by Thermo Fisher Scientific (1 mentions)
Cd25 pe cy7, by BD (1 mentions)
Cd11c pe cy7, by BioLegend (1 mentions)
Cd11c apc, by BioLegend (1 mentions)
3 protocols using mhc 2 pacific blue
1
Immune Cell Isolation and Analysis
2
Tracking Antigen-Presenting Cells in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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BALB/c mice (n=15/group) were injected subcutaneously in the footpad with 3×1010 vector genomes of ID-VP02 encoding GFP, control ID-VP02 encoding the cancer-testis antigen NY-ESO-1, or left untreated. Four days later, the draining popliteal or nondraining cervical lymph nodes were pooled from 5 mice (3 pools per treatment group) and live (L/D singlet events (L/D NIR−, SSH=SSA) were analyzed for the presence of GFP. The phenotype of GFP+ cells was determined by costaining with anti-mouse CD11c-PE-Cy7 (eBioscience), MHC II-Pacific Blue (eBioscience), SIGNR5-PE (eBioscience), and SIGNR1-APC (eBioscience).
3
Multicolor Flow Cytometry of Murine Immune Cells
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