Rat Interferon gamma ELISPOT Kit (ab64032)

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Overview

  • Product name

    Rat Interferon gamma ELISPOT Kit
    See all Interferon gamma kits

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Cell culture extracts

  • Assay type

    Sandwich (qualitative)

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Rat

  • Product overview

    The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.

    Abcam Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.

    Principal

    After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.

     

    Recognizes natural rat Interferon gamma.

  • Tested applications

    Suitable for: ELISpotmore details

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests 5 x 96 tests
    Bovine Serum Albumin 1 x 2g 1 x 2g
    IFNγ Biotinylated Detection Antibody 1 x 100µl 1 vial
    Rat IFNγ Pre-coated 96-well PDVF-bottomed plates 2 units 5 units
    Ready-to-use BCIP/NBT substrate buffer 1 x 11ml 2 x 25ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 10µl 1 x 50µl

  • Research areas

  • 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

  • Alternative names

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

  • Database links

    Applications

    Our Abpromise guarantee covers the use of ab64032 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
    ELISpot Use at an assay dependent dilution.

    References

    ab64032 has not yet been referenced specifically in any publications.

    Publishing research using ab64032? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Delivery plus Positive Control Issues

     Below average Good 4/5 (Ease of Use)
    Abreviews
    I ordered this product in July when Abcam's site said a 1-2 week lead time on delivery. I received the product roughly 4 weeks later after several emails apologizing for the product's delay.

    I unsuccessfully used the first 3 plates (following each step in the instruction book carefully) on freshly isolated rat PBMCs and splenocytes, without any spots on my positive controls of varying PMA + ionomycin concentrations. Abcam recommends PMA + ionomycin at 1 and 500 ng/mL in the instruction booklet, but PHA at 10 ug/ml here: https://www.abcam.com/protocols/elispot-protocol. I decided to try PHA at this concentration and my spots appeared beautifully, with nothing on the PMA+ionomycin wells.

    Between the delay and the instruction booklet setback, I have to give this product 2 stars.

    Benjamin Maimon

    Verified customer

    Submitted Oct 25 2017

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