Spleen cells were isolated and stained, as shown earlier (Farooque et al., 2014a (link)), following the lysis of red blood cells (RBCs) by ammonium chloride containing RBC lysis buffer (HiMedia). Splenocytes (1 × 106) or RAW 264.7 cells were re-suspended in 100 μL staining buffer containing fluorochrome-coupled antibodies: PE-labeled anti-mouse CD80, FITC-labeled anti-mouse CD86, PE-labeled anti-mouse MHCII and PECy5 anti-mouse F4/80 (eBioscience), and cells were incubated for 45 min in 4°C. Data were acquired on the BD LSR II and analyzed by using FlowJo software.
Pe labeled anti mouse mhcii
Manufactured by Thermo Fisher Scientific
The PE-labeled anti-mouse MHCII is a flow cytometry reagent used for the detection and quantification of major histocompatibility complex class II (MHCII) molecules on the surface of mouse cells. The PE fluorescent dye is conjugated to the anti-mouse MHCII antibody, enabling the specific labeling and identification of MHCII-positive cells in a sample.
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Lab products found in correlation
Lsr 2, by BD (1 mentions)
Rbc lysis buffer, by HiMedia (1 mentions)
Pe labeled anti mouse cd80, by Thermo Fisher Scientific (1 mentions)
Fitc labeled anti mouse cd86, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Pe labeled anti mouse cd11c, by BioLegend (1 mentions)
Fitc labeled anti mouse cd80, by BioLegend (1 mentions)
Mouse tnf α elisa kit, by Thermo Fisher Scientific (1 mentions)
Mouse il 6 elisa kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using pe labeled anti mouse mhcii
1
Flow Cytometry Phenotyping of Splenocytes
2
Murine Bone Marrow-Derived Dendritic Cell Isolation and Activation
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Bone marrow cells were isolated from C57BL/6 mice and cultured in RPMI 1640 media (GIBCO) supplemented with 10% heat inactivated FBS (BioInd), 1% penicillin/streptomycin (Beyotime), 20 ng/mL of IL-4 (Peprotech)and 20 ng/mL of GM-CSF (Peprotech) at 37 C and 5% CO 2 . Non-adherent cells were harvested on day 7 and used immediately. DC purity was confirmed with PE labeled antimouse CD11c (Biolegend) by FACS, consistently found to be > 90% CD11c + .
In vitro BMDC stimulation Immature BMDCs (5 3 10 4 ) were planted onto 96 well plates, then treated with simvastatin (1 mM) or LPS (100 ng/mL) for 24 h. The cell supernatants IL-6 was measured by mouse IL-6 ELISA kit (Ebioscience), TNF-a was measured by mouse TNF-a ELISA kit (Ebioscience) and IL-12p70 was measured by mouse IL-12p70 ELISA kit (Ebioscience) according to the manufacturer's instructions. Cell-surface co-stimulatory molecules and MHC-II were detected by FITC labeled anti-mouse CD80 (Biolegend), anti-mouse CD86 (Biolegend) and PE labeled anti-mouse MHC-II (Ebioscience).
In vitro BMDC stimulation Immature BMDCs (5 3 10 4 ) were planted onto 96 well plates, then treated with simvastatin (1 mM) or LPS (100 ng/mL) for 24 h. The cell supernatants IL-6 was measured by mouse IL-6 ELISA kit (Ebioscience), TNF-a was measured by mouse TNF-a ELISA kit (Ebioscience) and IL-12p70 was measured by mouse IL-12p70 ELISA kit (Ebioscience) according to the manufacturer's instructions. Cell-surface co-stimulatory molecules and MHC-II were detected by FITC labeled anti-mouse CD80 (Biolegend), anti-mouse CD86 (Biolegend) and PE labeled anti-mouse MHC-II (Ebioscience).
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