Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab25101 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
ELISA Use at an assay dependent concentration.
ELISpot Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration.

In vitro neutralization.  One Neutralizing Unit is defined as the total amount of antibodies sufficient for neutralizing one laboratory unit of recombinant Human IFNG (1 Unit = ~50 pg of pure Human IFNG).  Activity ≥ 10^4 Neutralizing U/mg protein.

ICC Use at an assay dependent concentration.

Follow an intracellular staining protocol.

WB Use at an assay dependent concentration. Predicted molecular weight: 19 kDa.
IHC-Fr Use at an assay dependent concentration.
Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [MD-1] to Interferon gamma (ab25014).

For sandwich ELISA, use this antibody as detection at 0.5µg/ml with ab25014 as capture.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • IFG antibody
    • IFI antibody
    • IFN gamma antibody
    • IFN, immune antibody
    • IFN-gamma antibody
    • IFNG antibody
    • IFNG_HUMAN antibody
    • Immune interferon antibody
    • Interferon gamma antibody
    see all

Images

  • ab25101 staining Human tonsil. Staining is localised to the cytoplasm and protein is secreted.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be

  • Standard curve for Interferon gamma (analyte: ab51240); dilution range 1pg/ml to 1µg/ml using capture antibody ab25014 at 5µg/ml and detector antibody ab25101 at 0.5µg/ml.

References

This product has been referenced in:
  • Zhang X  et al. Immunogenicity of adenovirus-vector vaccine targeting hepatitis B virus: non-clinical safety assessment in non-human primates. Virol J 15:111 (2018). Read more (PubMed: 30041659) »
  • Shen Y  et al. Metabolic control of the scaffold protein TKS5 in tissue-invasive, proinflammatory T cells. Nat Immunol 18:1025-1034 (2017). Read more (PubMed: 28737753) »
See all 4 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunohistochemistry (Resin sections)
Sample
Human Tissue sections (Tonsil)
Specification
Tonsil

Mr. Ali Hosseini

Verified customer

Submitted Oct 27 2014

Answer

Thank you for contacting us.

I have attached the filled out Exel spreadsheet with all the requested information. To be fair, all the information asked could be answered with a look on the datasheet...

In addition, I had a look through our tissue slices for her IHC-P experiments. Instead of the Tonsils she can use Lymphknot (as we do not have Tonsil slides).

https://www.abcam.com/index.html?datasheet=4350 (or use the following: https://www.abcam.com/index.html?datasheet=4350). PC for ab25101and ab10563

https://www.abcam.com/index.html?datasheet=4348 (or use the following: https://www.abcam.com/index.html?datasheet=4348). PC for ab34843

https://www.abcam.com/index.html?datasheet=4372 (or use the following: https://www.abcam.com/index.html?datasheet=4372). PC for ab37158

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Answer

We have a pair of anti-IFN gamma monoclonals that have been tested in sandwich ELISA:

Capture - ab8318

Detection - ab10076

Click here (or use the following: https://www.abcam.com/index.html?datasheet=8318).

Click here (or use the following: https://www.abcam.com/index.html?datasheet=10076).

I hope this helps. Please let me know if you have any questions.

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Answer

Thank you for your interest in our IFN gamma antibodies.

I recommend using your monoclonal antibody for capture in a sandwich ELISA, and a rabbit anti-human IFN gamma polyclonal antibody raised against full-length protein for detection, followed by an anti-rabbit IgG secondary antibody, pre-adsorbed against mouse IgG to remove reactivity with mouse (assuming your monoclonal antibody is mouse IgG). The secondary will be conjugated to HRP or AP or fluorochrome, depending on how you anticipate detecting the signal.

I suggest ab25101 for the polyclonal IFN gamma antibody,

Click here (or use the following: https://www.abcam.com/index.html?datasheet=25101)

and I suggest ab97080 for the anti-rabbit secondary, assuming you use HRP plus substrate for detection.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=97080).

The epitopes the antibody recognizes are not mapped, since this is a polyclonal antibody which recognizes multiple epitopes along the entire length of IFN gamma. If you want a monoclonal antibody (which recognizes a single epitope, rather than many) to pair with your monoclonal, I will need to know the epitope that your antibody recognizes in order to select an appropriate match. One of the two mouse monoclonal antibodies will need to be directly conjugated to an enzyme or fluorochrome, to avoid using an anti-mouse IgG secondary antibody.

I hope this is helpful. Please let me know if you have questions.

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Answer

Thank you for contacting us. We can certainly provide these in 500ug or 1mg size as a special request. Please let me know if you would like to place an order. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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