Human Granzyme B Quantikine HS ELISA Kit Summary
Sample Type & Volume Required Per Well
Serum (50 uL), Plasma (50 uL), Heparin Plasma (13 uL), Saliva (25 uL)
Assay Range
1.1 - 70 pg/mL (Serum, Plasma, Heparin Plasma, Saliva)
Specificity
Natural and recombinant human Granzyme B.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine® HS Human Granzyme B Immunoassay is a 4.0 hour solid-phase ELISA designed to measure human Granzyme B in serum, plasma and saliva. It contains NS0-expressed recombinant human Granzyme B and antibodies raised against the recombinant protein. Results obtained using natural human Granzyme B showed linear curves that were parallel to
the standard curves obtained using the Quantikine® kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Granzyme B.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians
Serum, Plasma, Heparin Plasma, Saliva
|
Intra-Assay Precision |
Inter-Assay Precision |
| Sample |
1 |
2 |
3 |
1 |
2 |
3 |
| n |
20 |
20 |
20 |
20 |
20 |
20 |
| Mean (pg/mL) |
10.5 |
16.5 |
42.8 |
10.8 |
17 |
42.9 |
| Standard Deviation |
0.117 |
0.267 |
0.566 |
0.627 |
0.83 |
2.02 |
| CV% |
1.1 |
1.6 |
1.3 |
5.8 |
4.9 |
4.7 |
Recovery
The recovery of human Granzyme B spiked to levels throughout the range of the assay was evaluated.
| Sample Type |
Average % Recovery |
Range % |
| EDTA Plasma (n=4) |
106 |
101-111 |
| Heparin Plasma (n=4) |
103 |
97-109 |
| Saliva (n=4) |
100 |
95-105 |
| Serum (n=4) |
108 |
101-115 |
Scientific Data
Human Granzybe B HS ELISA
Product Datasheets
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Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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).
Entrez Gene IDs:
3002 (Human); 14939 (Mouse)
Alternate Names:
C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; CTLA1; CTLA-1; CTSGL1; Fragmentin-2; Granzyme B; Granzyme-2; GranzymeB; GRB; GrzB; GZMB; HLP; SECT
