Recombinant
RabMAb

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

Overview

  • Product name

    Anti-Interferon alpha 2 antibody [EPR19074]
    See all Interferon alpha 2 primary antibodies
  • Description

    Rabbit monoclonal [EPR19074] to Interferon alpha 2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IP, ICCmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Interferon alpha 2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P01563

  • Positive control

    • WB: HEK-293T transfected with Interferon alpha 2 expression vector with a GFP-Myc-tag, whole cell lysate. ICC/IF: HEK-293T cells transfected with Interferon alpha 2 expression vector with a GFP-Myc-tag / GFP-tag. ICC: HEK-293T transfected with Interferon alpha 2 (pcDNA3.1 (+)-EGFP-Myc) IP: HEK-293T cells transfected with Interferon alpha 2 expression vector with a GFP-His-tag, whole cell lysate. Flow Cyt: HEK-293T cells transfected with Interferon alpha 2 expression vector with a GFP-His-tag
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19074
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab196221 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
WB 1/1000. Predicted molecular weight: 22 kDa.
Flow Cyt 1/600.
IP 1/30.
ICC 1/1000.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Target

  • Function

    Produced by macrophages, IFN-alpha have antiviral activities.
  • Sequence similarities

    Belongs to the alpha/beta interferon family.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • Alpha 2a interferon antibody
    • IFN alpha 2 antibody
    • IFN alphaA antibody
    • IFN-alpha-2 antibody
    • IFNA antibody
    • Ifna2 antibody
    • IFNA2_HUMAN antibody
    • IFNA2B antibody
    • INFA2 antibody
    • Interferon alpha 2a antibody
    • Interferon alpha 2b antibody
    • Interferon alpha A antibody
    • Interferon alpha-2 antibody
    • Interferon alpha-A antibody
    • Interferon antibody
    • LeIF A antibody
    • LeIFA antibody
    • MGC125764 antibody
    • MGC125765 antibody
    see all

Images

  • All lanes : 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
    Lane 2 : HEK-293T transfected with Interferon alpha 2 expression vector containing a GFP-Myc-tag, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 22 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    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.

  • 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.

  • 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.

  • 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.

  • 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.

References

ab196221 has not yet been referenced specifically in any publications.

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