Human Granzyme B ELISA Kit
Protocol Booklet
Key features and details
- Specification:
- Sensitivity:
- Standard Curve Range:
- Standard Curve Gradient:
- Number of Incubations:
- Sample Volume:
- Assay type:
- Operation Duration:
-
Brand:
CAT.NO. : AEH0052
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Product Details
Background
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.
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.
Typical data
| 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%.
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 in this assay. Donor medical histories were not available.
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