Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human IL-33 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling IL-33 with ab316846 at 1/50000 dilution (0.001 ug)/Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with Myc tag conjugated to Alexa Fluor®647.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling IL-33 with ab316846 at 1/500 (0.994 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear and cytoplasmic staining in HUVEC cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling IL-33 with ab316846 at 1/500 (0.994 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear and weak cytoplasmic staining in THP-1 treated with lipopolysaccharide (LPS, 5 ug/ml) and Phorbol-12-myristate-13-acetate (PMA, 50nM) for 24 hours.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HUVEC (human umbilical vein endothelial cell) treated with 500 pg/ml TNF-α for 24h and 300ng/ml BFA for 8h (Red) / Untreated HUVEC (Dotted Red) cells labelling IL-33 with ab316846 at 1/50 dilution (1 ug)/Red and Dotted Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

PMID : 18787100. Weaker staining occurred in HUVEC cells treated with 500 pg/ml TNF-α for 24h and 300ng/ml BFA for 8h compared to untreated HUVEC cells.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte) treated with 5ug/ml LPS and 50nM PMA for 24h (Red) / Untreated THP-1 (Dotted Red) cells labelling IL-33 with ab316846 at 1/50 dilution (1 ug)/Red and Dotted Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Stonger staining occurred in THP-1 treated with 5ug/ml LPS and 50nM PMA for 24h compared to untreated THP-1 cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-33 antibody [EPR26367-2] - BSA and Azide free (AB316847)

This data was developed using ab316846, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial) cells labelling IL-33 with ab316846 at 1/500 dilution, followed by Goat anti-Rabbit (AlexaFluor^® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing the nuclear and weak cytoplasmic staining (shown in green) is decreased after the treatment with IL-1β (5 ng/ml) for 24 hours in HUVEC cells. Nuclear DNA was labeled with DAPI (shown in blue).

Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A]-Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (shown in magenta).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

The reduction in IL-33 when stimulated with TNFa in HUVEC cells (specially the nuclear IL-33) (PMID : 18787100).