Human Granzyme B ELISA Assay


The Human Granzyme B ELISA Assay is to be used for the in-vitro quantitative determination of Granzyme B in human serum, plasma, buffered solutions or cell culture medium. The Eagle Biosciences Human Granzyme B ELISA Assay kit is for Research Use Only.

SKU: GRB31-K01 Categories: , ,

Human Granzyme B ELISA Assay

The Human Granzyme B ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 20 pg /ml
Dynamic Range: 31.25 pg/ml – 1000 pg/ml
Incubation Time: 3h
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl

Assay Background

Granzyme B belongs to the granzymes family, which are exogenous serine proteinases. Granzymes are produced by cytotoxic lymphocytes (CTLs) and NK cells.

Activated cytotoxic T lymphocytes and NK cells are important effectors in immunologic defenses against tumor cell proliferation, viral infection and transplant tissue. They induce apoptosis via multiple mechanism thanks to granules exocytosis. Following adhesion of the CTL on the target cell (via CTL-receptor and antigen-presenting MHC molecules), CTLs secrete cytoplasmic granules containing granules and enzymes including granzymes and perforine (pore-forming protein). Granules are then released in the intercellular space, perforine forms a transmembrane pore. Through these pores, granzymes enter the cytosol of the target cell.

Granzyme B is the most powerful pro-apoptotic member of granzyme family. Granzyme B activates the intracellular caspases induising finally the death of the cell.

Granzymes are diagnostics markers for many disorders (viral infection, lymphoma’s and carcinoma’s, auto-immune disease and rejection of transplant).

Related Products

Human Perforin ELISA Assay Kit
Human MICA ELISA Assay Kit
Human CD138 /SYNDECAN-1 ELISA Assay Kit

Additional Information

Assay Principle

The Granzyme B Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for Granzyme B has been coated onto the wells of the microtiter strips provided. Samples, including standards of known Granzyme B concentrations and unknowns are pipetted into these wells.  During the first incubation, the Granzyme B antigen and a biotinylated monoclonal antibody specific for Granzyme B are simultaneously incubated.

After washing, the enzyme ( streptavidin-peroxydase ) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution which is acting on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of Granzyme B present in the samples.

Assay Procedure

  1. Add 100µl of each standard, sample and zero in duplicate to appropriate number of wells
  2. Add 50µl of diluted biotinylated anti-Granzyme B to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 3 hours
  4. Remove the cover and wash the plate
  5. Add 100μl of Streptavidin-HRP solution into all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  7. Repeat wash step 5.
  8. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  9. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  10. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


Product Documents

Product Citations