Anti-Interferon alpha 2 antibody [EPR19074] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Interferon alpha 2 antibody [EPR19074] - BSA and Azide free (AB230830)

Intracellular flow cytometric analysis of�4% 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 Fluor^� 647) (ab150079), at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196221).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Interferon alpha 2 antibody [EPR19074] - BSA and Azide free (AB230830)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196221).

• ICC

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Immunocytochemistry - Anti-Interferon alpha 2 antibody [EPR19074] - BSA and Azide free (AB230830)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196221).

• IP

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Immunoprecipitation - Anti-Interferon alpha 2 antibody [EPR19074] - BSA and Azide free (AB230830)

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 as for detection at 1/10000 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196221).

All lanes:

Immunoprecipitation - Anti-Interferon alpha 2 antibody [EPR19074] (<a href='/en-us/products/primary-antibodies/interferon-alpha-2-antibody-epr19074-ab196221'>ab196221</a>)

Predicted band size: 22 kDa

Observed band size: 50 kDa

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