Human Granzyme B enzyme-linked immunoassay kit
| Specification | 96 Test |
|---|---|
| Sensitivity | 6.62 pg/ml (50 μl) |
| Standard Curve Range | 78.13~5000 pg/ml |
| Standard Curve Gradient | 7 Points/3 Folds |
| Number of Incubations | 2 |
| Detectable sample | Liquid phase sample of soluble substances. For example: serum, plasma, cell culture supernatant, tissue grinding liquid, etc. |
| Sample Volume | 50 μl/10 μl |
| Type | Full ready-to-Use |
| Operation Duration | 120min |
| pg/ml | O.D. | Average | Corrected | |
|---|---|---|---|---|
| 0.00 | 0.0354 | 0.0305 | 0.0330 | |
| 78.13 | 0.0862 | 0.0936 | 0.0899 | 0.0570 |
| 156.25 | 0.1339 | 0.1493 | 0.1416 | 0.1087 |
| 312.50 | 0.2451 | 0.2431 | 0.2441 | 0.2112 |
| 625.00 | 0.3759 | 0.3953 | 0.3856 | 0.3527 |
| 1250.00 | 0.7970 | 0.7870 | 0.7920 | 0.7591 |
| 2500.00 | 1.4500 | 1.3650 | 1.4075 | 1.3746 |
| 5000.00 | 2.4690 | 2.3120 | 2.3905 | 2.3576 |
Precision
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample Number | S1 | S2 | S3 | S1 | S2 | S3 |
| 22 | 22 | 22 | 6 | 6 | 6 | |
| Average(pg/ml) | 97.1 | 448.0 | 1618.5 | 84.9 | 453.4 | 1514.2 |
| Standard Deviation | 5.1 | 17.4 | 58.5 | 3.7 | 10.0 | 31.0 |
| Coefficient of Variation(%) | 5.2 | 3.9 | 3.6 | 4.3 | 2.2 | 2.1 |
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 human Granzyme B into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 101% to 128% with an overall mean recovery of 111%.
Sample Values
| Sample Matrix | Sample Evaluated | Range (pg/ml) | Detectable (%) | Mean of Detectable (pg/ml) |
|---|---|---|---|---|
| Serum | 30 | n.d.-58.36 | 37.5 | 14.21 |
Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of Granzyme B in this assay. No medical histories were available for the donors.
Product Data Sheet
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.
