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Interferon Gamma + Granzyme B Human Elispot Kit (Reagents for 15 x 96 Well Plates) (ab48763)

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Overview

Product name

Interferon Gamma + Granzyme B Human Elispot Kit (Reagents for 15 x 96 Well Plates)
See all Interferon Gamma + Granzyme B products (12) ...

Tests

15 x 96 well plate

Sample type

Cell culture supernatant, Cell culture extracts

Assay type

ELISPOT

Cross reactivity

Reacts with

Human

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, oncology, infectious diseases, autoimmune diseases and transplantation.
The 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.

The dual colour Elispot allows you to monitor the production of two cytokines simultaneously in the same well.



Principle



After cell stimulation, locally produced cytokines are captured by Interferon gamma and Granzyme B specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti- Interferon gamma FITC conjugated antibody and a biotinylated anti- Granzyme B antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate Interferon gamma production while Granzyme B is revealed by blue/purple spots.


Tested applications

Measuring secreted proteins from cellsmore details

Properties

Storage instructions

Store at 4°C. The kit is shipped at room temperature

Components 15 x 96 tests
10 x concentrate buffer for the preparation of AEC buffer 3 x 5ml
50 x concentrate AEC substrate buffer 3 x 1ml
Anti-FITC antibody HRP conjugate 3 x 100µl
Biotinylated detection antibody for Granzyme B (resuspend each in 0.55mL)3 x 0.55ml
Bovine Serum albumin 3 x 1g
Capture antibody for Granzyme B 3 x 0.5ml
Capture antibody for Interferon gamma 3 x 0.5ml
Dry skimmed milk 3 x 1g
FITC conjugated detection antibody for Interferon gamma (resuspend each in 0.55mL)3 x 0.55ml
Ready-to-use BCIP/NBT substrate buffer3 x 50ml
Streptavidin Alkaline Phosphatase conjugate 3 x 50µl

Research areas

Kits/ Lysates/ Other >> Kits >> ELISPOT and FLUOROSPOT Kits >> ELISPOT Kits >> Interferon ELISPOT kits
Kits/ Lysates/ Other >> Kits >> ELISPOT and FLUOROSPOT Kits >> FLUOROSPOT Kits >> Interferon FLUOROSPOT kits
Cancer >> Tumor immunology >> Cytokines >> Interferons
Stem Cells >> Signaling Pathways >> TGF beta >> Interferon gamma
Immunology >> Innate Immunity >> Cytokines >> Interferons
Cell Biology >> Apoptosis >> Extracellular Signals >> Granzymes

Relevance

Interferon Gamma is 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. Granzyme B is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis.

Alternative names

  • CSPB
  • Cytotoxic T lymphocyte proteinase 2
  • Granzyme 2
  • Granzyme B
  • GZMB
  • HLP
  • IFI
  • IFN gamma
  • IFNG
  • Immune interferon
see all

Database links

Applications

Show applications key

Our Abpromise guarantee covers the use of ab48763 in the following tested applications.

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

  • ELISPOT

     ELISPOT

    ELISPOT

Protocols

  • Human Interferon gamma + Granzyme B Elispot Kit

    • References for Interferon Gamma + Granzyme B Human Elispot Kit (Reagents for 15 x 96 Well Plates) (ab48763)

    ab48763 has not yet been referenced specifically in any publications.

    Publishing research using ab48763? Please let us know so that we can cite the reference in this datasheet

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