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.
Mhcii 1 a 1 e pacific blue
Manufactured by BioLegend
The MHCII (I-A/I-E)-Pacific Blue is a fluorophore-conjugated antibody that binds to the major histocompatibility complex class II (MHCII) molecules, specifically the I-A and I-E regions, in mouse cells. This product can be used for the detection and analysis of MHCII-expressing cells through flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cd11b alexa fluor 700, by BD (1 mentions)
Cd11c pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd8α apc h7, by BD (1 mentions)
Cd19 apc h7, by BD (1 mentions)
Cd19 bv510, by BD (1 mentions)
Mhc 2 pacific blue, by Thermo Fisher Scientific (1 mentions)
Cd138 pe, by BioLegend (1 mentions)
Thy1.1 percp, by BioLegend (1 mentions)
Flojo software, by Tree Star (1 mentions)
Cd4 apc, by BioLegend (1 mentions)
Percoll, by GE Healthcare (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Cd45 fitc, by BD (1 mentions)
Cd86 pe, by BioLegend (1 mentions)
Cd11c pe cy7, by BioLegend (1 mentions)
Ccr7 apc, by BioLegend (1 mentions)
Cd45 percp cy5, by BioLegend (1 mentions)
Lsrfortessa cell analyzer, by BD (1 mentions)
2 protocols using mhcii 1 a 1 e pacific blue
1
Immune Cell Isolation and Analysis
2
Immunophenotyping of Lymph Node and Lung Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To determine the composition and activation of immune cells in the lymph nodes from vaccinated mice and lungs from infected mice, single-cell suspension were prepared and stained with fluorochrome-conjugated antibodies before fixing in 4% paraformaldehyde for flow cytometry assay. Single cells from lymph nodes were stained with CD45-PerCP/Cy5.5, B220-Brilliant Violet 421, CD11c-PE/Cy7, CCR7-APC, CD86-PE, and MHC-II (I-A/I-E)-Pacific blue antibodies (all from Biolegend) for the identification the activation of dendritic and B cells. Single cells from lung tissues were stained with CD45-APC/Cy7, B220-PerCP/Cy5.5, CD3-PerCP/Cy5.5, CD4-PE/Cy7, CD8-PE, CD44-APC/Cy7, and CD62L-Brilliant Violet 421 antibodies. Effector memory T cells were identified as CD3+, CD4+/CD8+, CD44+, and CD62L− cells, whereas central memory T cells were identified as CD3+, CD4+/CD8+, CD44+, and CD62L+ cells. Stained cells were analyzed on LSRFortessa Cell Analyzer (BD Bioscience), data were analyzed using FlowJo software (TreeStar, Inc.).
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