Anti-IFITM2+IFITM3 antibody [EPR28942-88]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 10ng/mL IFN-alpha 1 for 16 hours (Green) / Untreated HeLa (Magenta) cells labelling IFITM2+IFITM3 with ab323747 at 1/50 dilution (1 ug) / Magenta and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling IFITM2+IFITM3 with ab323747 at 1/250 (2.004 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in HeLa cell line treated with IFN alpha 1 (10ng/ml) for 16h, and weak cytoplasmic staining in HeLa cell line (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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml 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 2ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell) treated with 10ng/mL IFN-alpha 1 for 16 hours (Green) / Untreated A549 (Magenta) cells labelling IFITM2+IFITM3 with ab323747 at 1/50 dilution (1 ug) / Magenta and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (upper left) / 293T (human embryonic kidney epithelial cell) cells transfected with an empty expression vector containing a Myc tag (upper middle) / 293T cells transfected with a IFITM1 expression vector containing a GFP tag (down left) / 293T cells transfected with a IFITM2 expression vector containing a GFP tag (down middle) / 293T cells transfected with a IFITM3 expression vector containing a GFP tag(down right) cells labelling IFITM2+IFITM3 with ab323747 at 1/5000 dilution (0.01 ug) 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling IFITM2+IFITM3 with ab323747 at 1/250 (2.004 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ 1000 2ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in A549 cell line treated with IFN alpha (1000 U/ml) for 24h, and weak staining in A549 cell line (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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml 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 2ug/ml dilution.

• WB

Supplier Data

Western blot - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34078739; PMID : 34321474; PMID : 30643282).

The expression of IFITM2 and IFITM3 is up-regulated in response to IFN alpha 1 or IFN beta treatment (PMID : 34078739; PMID : 34321474; PMID : 30643282).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 15 seconds; Lanes 5-6 : 5 seconds

All lanes:

Western blot - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (ab323747) at 1/1000 dilution

Lanes 1 and 3:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 treated with 1000U/ml IFN alpha 1 for 24 hours whole cell lysate at 20 µg

Lane 4:

A549 treated with 1000U/ml IFN beta 1 for 24 hours whole cell lysate at 20 µg

Lane 5:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HeLa treated with 10ng/ml IFN alpha 1 for 16 hours 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 1/20000 dilution

Observed band size: 15 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (AB323747)

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

Negative control : Daudi

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34078739; PMID : 34321474; PMID : 30643282).

Lane 3 is incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 1-2 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IFITM2+IFITM3 antibody [EPR28942-88] (ab323747) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

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 1/20000 dilution

Lane 3:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 15 kDa,36 kDa

false

Exposure time: 70s