Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
39.1-2500 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Human Granzyme B DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Background: Granzyme B
Granzyme B is a member of the granzyme family of serine proteases found specifically in
granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells (1, 2). Granzyme B plays
an essential role in granule-mediated apoptosis utilizing the substrates in this pathway, such as
Caspase 3, Caspase 8 and Bid (3, 4). Recent research indicates expanded Granzyme B
functionality to include extracellular roles along with its classical pro-apoptotic function. It has
been found that Granzyme B is an important mediator of skin injury, repair and inflammation
(4) through extracellular substrates including Laminin, VE-Cadherin, Fibronectin and the
proteoglycans Aggrecan (3) and Decorin (4). As one of the five Granzymes (A, B, H, K and M) identified in the human genome, Granzyme B
(32kDa) (5) is the most widely researched in terms of its biological function and its utility in
health and disease (4). It is synthesized as a precursor (247 residues) with a signal peptide
(residues 1-18), a pro-peptide (residues 19-20), and a mature chain (residues 21-247) (6-8).
Once inside granules, Granzyme B is fully processed into the mature chain and becomes an
active protease when the pro-peptide, Gly-Glu is removed from the N-terminus by cleavage
with Cathepsin C (9). The protease activity of Granzyme B is tightly controlled by Serpin B9/
Protease Inhibitor 9 (9). The amino acid sequence of human Granzyme B is 71%, 69%, and 68%
identical to its canine, rat, and mouse counterparts, respectively.
Granzymes have been shown to modulate inflammation, and Granzyme B plasma levels have
been found higher with atopic dermatitis and psoriasis when compared to healthy controls.
This is in contrast to Granzyme A plasma levels which remain unchanged (10). Serum from
patients with Crohn's disease have significantly higher Granzyme B levels than controls (11).
Alternate Names
CGL-1, CGL1, CSPB, CTLA-1, CTLA1, CTSGL1, Fragmentin-2, Granzyme-2, GRB, GrzB, GZMB, HLP, SECT
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