Intended use

This immunoassay kit allows for the specific measurement of Rat granzymes B concentrations in cell culture supernates, serum and plasma.

 

Introduction

Granzyme B is a member of the Granzyme family of serine proteases which are foundspecifically in the cytotoxic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK)cells. As the two major populations of cytotoxic lymphocytes (CL), CTL and NK cells are key components of the innate and adaptive cellular immune responses against intracellular pathogens and transformed cells. Released through exocytosis of CL secretory granule contents and aided by perforin, Granzyme B is able to access the target cell cytosol, where it processes key substrates to trigger cell death. In addition to playing an essential role in granule-mediated apoptosis, Granzyme B may have roles in rheumatoid arthritis and in bacterial and viral infection. The substrates that have been identified for Granzyme B include caspases, BID, lamins, poly(ADP-ribose) polymerase, neuronal glutamate receptor and cartilage proteoglycan.

Granzyme B is the most powerful pro-apoptotic member of the granzyme family. Like caspases, cysteine proteases that play an important role in apoptosis, it can cleave proteins after acidic residues, especially aspartic acid.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Granzymes B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Granzymes B present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for Granzymes B is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of Granzymes B bound in the initial step. The color development is stopped and the intensity of the color is measured.