Mouse Granzyme B enzyme-linked immunoassay kit

CAT: EM0016 Datasheet
Specification 96 Test
Sensitivity 9.91 pg/ml (10 μl)
Standard Curve Range 27.43~20000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Sample Volume 10 μl
Type Fully Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0150 0.0130 0.0140
27.43 0.0230 0.0200 0.0215 0.0075
82.30 0.0340 0.0290 0.0315 0.0175
246.91 0.0680 0.0670 0.0675 0.0535
740.74 0.1650 0.1630 0.1640 0.1500
2222.22 0.4570 0.4220 0.4395 0.4255
6666.67 1.2100 1.1460 1.1780 1.1640
20000.00 2.7790 2.7510 2.7650 2.7510

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 341.6 1646.5 5204.5 367.9 1876.2 5851.4
Standard Deviation 22.6 94.6 295.4 8.6 63.2 192.3
Coefficient of Variation(%) 6.6 5.7 5.7 2.3 3.4 3.3

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 six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of Mouse Granzyme B into health mouse serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 107% to 132% with an overall mean recovery of 116%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 10.64-35.31 92 24.13

Serum/Plasma – Thirty samples from apparently healthy mice were evaluated for the presence of Granzyme B in this assay. No medical histories were available for the donors. n.d. = non-detectable. Samples measured below the sensitivity are considered to be non-detectable.

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. Granzyme B plays an essential role in granule-mediated apoptosis utilizing the substrates in this pathway, such as Caspase 3, Caspase 8 and Bid. 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 through extracellular substrates including Laminin, VE-Cadherin, Fibronectin and the proteoglycans Aggrecan and Decorin.

As one of the five Granzymes (A, B, H, K and M) identified in the human genome, Granzyme B (32kDa) is the most widely researched in terms of its biological function and its utility in health and disease . 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). 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. The protease activity of Granzyme B is tightly controlled by Serpin B9/ Protease Inhibitor 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. Serum from patients with Crohn's disease have significantly higher Granzyme B levels than controls.

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