Cell Biology
Mast Cell Degranulation Assay Kit
- Analyze tryptase activity in cell lysates, supernatants
- Tryptase inhibitor and inducer included for screening purposes
- pNA Standard is provided as a comparison control
Mast cells are highly granulated cells found in connective tissues at maturity, and are important regulators of various inflammation conditions, in particular, allergic reactions. Allergic reactions result from the release of proinflammatory mediators from the mast cells, among them heparin, histamine, and proteases such as chymase and tryptase. A tetrameric serine protease, tryptase has emerged as the major component of mast cell granules, comprising up to 20% of the total protein of mast cells derived from lung, colon and skin tissue. The characteristic feature of tryptase release from mast cells is a representative indicator of mast cell activation during an inflammatory response, and because tryptase is stored almost exclusively in mast cells, it is a target for therapeutic intervention in allergic diseases.
Tryptase Activity of Basophilic Cell Lysates Human KU812 and Rat RBL basophils are treated with Calcium Ionophore A23187 (+/-Protamine). After a one hour incubation period at 37 C, cells are lysed, and tryptase activity is determined on a standard microplate reader. |
The Mast Cell Degranulation Assay Kit is a live cell assay kit that offers an efficient and sensitive system for the evaluation of tryptase activity in cell lysates, supernatants or for inhibitor screening. The assay is based on spectophotometric detection of free chromophore p-nitroaniline (pNA) after it is cleaved from the labeled tryptase substrate tosyl-gly-pro-lys-pNA.
Kit Components
- Tryptase Positive Control
- Assay Buffer
- Calcium Ionophore
- Tryptase Inhibitor (Protamine)
- Tryptase Substrate (tosyl-gly-pro-lys-pNA)
- pNA Standard
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