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    products/primary-antibodies/interferon-gamma-antibody-epr23991-53-ab267369.pdf

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Immunology Innate Immunity Cytokines Interferons
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Interferon gamma antibody [EPR23991-53] (ab267369)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question

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Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
  • Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
  • Flow Cytometry (Intracellular) - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
  • Anti-Interferon gamma antibody [EPR23991-53] (ab267369)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23991-53] to Interferon gamma
  • Suitable for: Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

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Knockout
Product image
Human IFNG knockout Jurkat cell line (ab273746)

View more associated products

Overview

  • Product name

    Anti-Interferon gamma antibody [EPR23991-53]
    See all Interferon gamma primary antibodies
  • Description

    Rabbit monoclonal [EPR23991-53] to Interferon gamma
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 ug/mL, 6 h), BFA (5 ug/ml, last 5 h) cell lysate. Flow Cyt (intra): PBMC cells treated with cell stimulation cocktail (80nM PMA+1.34µM Ionomycin+10.6µM BFA+2uM Monensin).
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23991-53
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interferons
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Interferon gamma
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interferons
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Interferon gamma antibody [EPR23991-53] - BSA and Azide free (ab280574)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Dyes/Markers

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab267369 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 19 kDa.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 19 kDa.
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
  • Tissue specificity

    Released primarily from activated T lymphocytes.
  • Involvement in disease

    In Caucasians, genetic variation in IFNG is associated with the risk of aplastic anemia (AA) [MIM:609135]. AA is a rare disease in which the reduction of the circulating blood cells results from damage to the stem cell pool in bone marrow. In most patients, the stem cell lesion is caused by an autoimmune attack. T-lymphocytes, activated by an endogenous or exogenous, and most often unknown antigenic stimulus, secrete cytokines, including IFN-gamma, which would in turn be able to suppress hematopoiesis.
  • Sequence similarities

    Belongs to the type II (or gamma) interferon family.
  • Post-translational
    modifications

    Proteolytic processing produces C-terminal heterogeneity, with proteins ending alternatively at Gly-150, Met-157 or Gly-161.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P01579 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3458 Human
    • Omim: 147570 Human
    • SwissProt: P01579 Human
    • Unigene: 856 Human
    • Alternative names

      • IF 1 antibody
      • IFG antibody
      • IFI antibody
      • ifn g antibody
      • IFN gamma antibody
      • IFN immune antibody
      • IFN, immune antibody
      • IFN-gamma antibody
      • IFNG antibody
      • IFNG_HUMAN antibody
      • Immune interferon antibody
      • Interferon gamma antibody
      • Type II Interferon antibody
      see all

    Images

    • Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      All lanes : Anti-Interferon gamma antibody [EPR23991-53] (ab267369) at 1/1000 dilution

      Lane 1 : Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg
      Lane 2 : IFNG knockout Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg
      Lane 3 : PTA-6967 Vehicle control + Brefeldin A (5 ug/ml, 6 h) cell lysate at 10 µg
      Lane 4 : PTA-6967 Treated TPA (80 nM, 5 h), Ionomycin ab120116 (3 µM, 5 h) + Brefeldin A (5 µg/mL, 3 h) cell lysate at 10 µg

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 28 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      Western blot - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      All lanes : Anti-Interferon gamma antibody [EPR23991-53] (ab267369) at 1/1000 dilution

      Lane 1 : Wild-type Jurkat Vehicle control: PMA (0 ng/mL, 6 h) and ionomycin (0 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 20 µg
      Lane 2 : Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 g/ml, last 5 h) cell lysate at 20 µg
      Lane 3 : IFNG knockout Jurkat Vehicle control: PMA (0 ng/mL, 6 h) and ionomycin (0 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 20 µg
      Lane 4 : IFNG knockout Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg

      Performed under reducing conditions.

      Predicted band size: 19 kDa
      Observed band size: 28 kDa why is the actual band size different from the predicted?



      Western blot: Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in treated wild-type Jurkat cell lysates with no signal observed at this size in treated IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) secondary antibody and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. Blot was developed with an ultra-high sensitivity ECL reagent.

    • Flow Cytometry (Intracellular) - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      Flow Cytometry (Intracellular) - Anti-Interferon gamma antibody [EPR23991-53] (ab267369)

      Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) treated with cell stimulation cocktail (80nM PMA+1.34uM Ionomycin+10.6uM Brefeldin A+2uM Monensin) for 6 hours cells labelling Interferon gamma with ab267369 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were stained with anti-CD4 conjugated to Pacific blue. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab267369.

    • Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
      Anti-Interferon gamma antibody [EPR23991-53] (ab267369)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab267369? Please let us know so that we can cite the reference in this datasheet.

    ab267369 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Interferon gamma antibody [EPR23991-53]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (RAW cells - murine macrophages)
    Permeabilization
    Yes - 0.2% Triton X-100 in PBS
    Specification
    RAW cells - murine macrophages
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Armen Petrosyan

    Verified customer

    Submitted Dec 03 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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