Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin (AB327881)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) and WEHI-231 (mouse B cell lymphoma B lymphocyte) labelling IL-6R with ab327881 at 1/100 (10.42 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membranous and cytoplasmic staining in RAW 264.7 cells and no staining in WEHI-231 cells(shown in green). The counterstain was observed in magenta.
Low expression : WEHI-231.
Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• Flow Cyt

Lab

Flow Cytometry - Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin (AB327881)

Flow cytometric analysis of Mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labelling IL-6R with ab327881 at 1/100 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Cells were co-stained with anti-CD19 conjugated to PE/CY7.

• Flow Cyt

Lab

Flow Cytometry - Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin (AB327881)

Flow cytometric analysis of Mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labelling IL-6R with ab327881 at 1/100 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Cells were co-stained with anti-CD4 conjugated to Alexa Fluor®488.

• Flow Cyt

Lab

Flow Cytometry - Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin (AB327881)

Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) and WEHI-231 (mouse B cell lymphoma B lymphocyte, Left) labelling IL-6R with ab327881 at 1/100 dilution (1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : WEHI-231.

• FuncS

Lab

Functional Studies - Anti-IL-6R antibody [EPR30382-502] - BSA and Azide free, low endotoxin (AB327881)

Proliferation inhibition of B9 cells following stimulation with mouse IL-6 in the presence of either an anti-IL6RA antibody (ab327881) or an IgG isotype control (ab199376).(A) Dose–response curve of cell proliferation of B9 cells exposed to increasing concentrations of mouse IL-6.(B) Proliferation inhibition induced by 100 ng/mL mouse IL-6 following treatment with anti-IL6RA antibody (circles, solid line) or IgG isotype control (triangles, dotted line) across a range of concentrations. The IC₅₀ for inhibition of proliferation is 0.41 µg/mL.B9 cells were seeded at 3 × 10⁴ cells per well in a 96-well plate. Cells were pre-incubated with a dilution series of anti-IL6RA antibody for 6 hours prior to IL-6 stimulation and then cultured for 48 hours in medium containing 5% FBS. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.