Anti-IFNL3 antibody [EPR30983-3]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IFNL3 antibody [EPR30983-3] (AB327709)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a human IFNL3 expression vector containing a HA-Myc-tag® (Upper) 293T cells transfected with a human IFNL2 expression vector containing a HA-Myc-tag® (Lower left) 293T cells transfected with an empty expression vector containing a myc-His-tag® (Lower right) cells labelling with ab327709 at 1/5000 dilution (0.01ug) / Upper right and Lower 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® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFNL3 antibody [EPR30983-3] (AB327709)

Immunohistochemical analysis of paraffin-embedded (A) : HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human IFNL3 expression vector containing a his tag. (B) : HEK-293T cells transfected with a human IFNL2 expression vector containing a his tag. (C) : HEK-293T cells transfected with empty vector containing a his tag. tissue labeling with ab327709 at 1/2000 (0.253 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T cells transfected with a human IFNL3 expression vector containing a his tag, no staining on (B) HEK-293T cells transfected with a human IFNL2 expression vector containing a his tag, and (C) HEK-293T cells transfected with empty vector containing a his tag.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IFNL3 antibody [EPR30983-3] (AB327709)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T cells transfected with an empty expression vector containing a myc-His-tag® HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human IFNL3 expression vector containing a His-Myc tag HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human IFNL2 expression vector containing a His-Myc tag cells labelling with ab327709 at 1/1000 (0.505 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 positive staining in HEK-293T transfected with a human IFNL3 expression vector containing a His-Myc tag(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (Magenta). 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 dilution.

• WB

Lab

Western blot - Anti-IFNL3 antibody [EPR30983-3] (AB327709)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with overexpressed human IFNL2 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (21KDa);.

All lanes:

Western blot - Anti-IFNL3 antibody [EPR30983-3] (ab327709) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human IFNL3 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human IFNL2 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 21 kDa,36 kDa

false

Exposure time: 26s