Anti-Interferon alpha 2 antibody [EPR19074]

• WB

Supplier Data

Western blot - Anti-Interferon alpha 2 antibody [EPR19074] (AB196221)

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular mass observed is due to the presence of the GFP epitope tag on full length IFN alpha 2.

All lanes:

Western blot - Anti-Interferon alpha 2 antibody [EPR19074] (ab196221) at 1/5000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty vector, containing a GFP-Myc-tag, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with Interferon alpha 2 expression vector containing a GFP-Myc-tag, whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 22 kDa

Observed band size: 50 kDa

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Exposure time: 3s

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Interferon alpha 2 antibody [EPR19074] (AB196221)

Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with Interferon alpha 2 expression vector containing a GFP-His-tag cell line) cells labeling Interferon alpha 2 with ab196221 at 1/600 (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluorr^® 647) (ab150079), at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Interferon alpha 2 antibody [EPR19074] (AB196221)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with either GFP-tagged Interferon alpha 2 expression vector or empty GFP expression vector) cells labeling Interferon alpha 2 with ab196221 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 647) (ab150079) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in HEK-293T cells transfected with GFP-tagged Interferon alpha 2 expression vector.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of ab196221, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 647) (ab150079) secondary antibody at 1/1000 dilution.

• ICC

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Immunocytochemistry - Anti-Interferon alpha 2 antibody [EPR19074] (AB196221)

Immunocytochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with Interferon alpha 2 (pcDNA3.1 (+)-EGFP-Myc)) cells labeling Interferon alpha 2 with ab196221 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use secondary antibody. Positive staining on HEK-293T cells transfected with Interferon alpha 2 (pcDNA3.1 (+)-EGFP-Myc) (image A); and no staining on HEK-293T cells without transfection (image B). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of ab196221, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.

• IP

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Immunoprecipitation - Anti-Interferon alpha 2 antibody [EPR19074] (AB196221)

Interferon alpha 2 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with Interferon alpha 2 expression vector containing a GFP-His-tag) whole cell lysate using ab196221 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab196221 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : HEK-293T (transfected with Interferon alpha 2 expression vector containing a GFP-His-tag) whole cell lysate 10 μg (Input).

Lane 2 : ab196221 IP in HEK-293T (transfected with Interferon alpha 2 expression vector containing a GFP-His-tag) whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab196221 inHEK-293T (transfected with Interferon alpha 2 expression vector containing a GFP-His-tag) whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

The molecular mass observed is due to the presence of the GFP epitope tag on full length IFN alpha 2.

All lanes:

Immunoprecipitation - Anti-Interferon alpha 2 antibody [EPR19074] (ab196221)

Predicted band size: 22 kDa

Observed band size: 50 kDa

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