Human Granzyme B enzyme-linked immunoassay kit

CAT: EH0052 Datasheet
Specification 96 Test
Sensitivity 1.12 pg/ml (50 μl);5.96 pg/ml (10 μl)
Standard Curve Range 6.86~5000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Sample Volume 50 μl/10 μl
Type Full ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0147 0.0140 0.0144
6.86 0.0275 0.0263 0.0269 0.0126
20.58 0.0453 0.0453 0.0453 0.0310
61.73 0.1044 0.1023 0.1034 0.0890
185.19 0.2562 0.2565 0.2564 0.2420
555.56 0.7001 0.7000 0.7001 0.6857
1666.67 1.6770 1.6800 1.6785 1.6642
5000.00 3.0660 3.0670 3.0665 3.0522

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 102% to 112% with an overall mean recovery of 110%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 9.62-835.77 100 230.34

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of IL-8 in this assay. No medical histories were available for the donors.

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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