MAST CELL DEGRANULATION ASSAY KIT

MAST CELL DEGRANULATION ASSAY KIT

Stock Code: 3587014
Manufacturer Part No: IMM001
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Application


The Mast Cell Degranulation Assay Kit provides a quick, efficient and sensitive system for evaluation of tryptase activity in culture supernatants, cell lysates or other tryptase-containing samples. Testing of purified tryptase enzyme, in vitro inhibitor screening and the study of cell degranulation can also be performed with this assay.

The Chemicon Mast Cell Degranulation Assay Kit is intended for research use only, and not for diagnostic or therapeutic applications.


The Mast Cell Degranulation Assay Kit provides a quick, efficient & sensitive system for evaluation of tryptase activity in culture supernatants, cell lysates or other tryptase-containing samples.


Components


Tryptase Positive Control (Part No. 2003650): 20 µL of 0.1 mg/mL solution.5X Assay Buffer (Part No. 2003649): 20 mL containing 0.1% heparin.Calcium Ionophore A23187 (Part No. 2003652): 100 µL of 1 mM in DMSO.Tryptase Inhibitor (Protamine) (Part No. 2003651): 250 µL of 10 mM solution.Tryptase Substrate (tosyl-gly-pro-lys-pNA) (Part No. 90569): 3.125 mg.pNA Standard (Part No. 90085): 250 µL of 10 mM in DMSO.


Disclaimer


Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.


General description


Introduction

Degranulation, the secretion of cytoplasmic granules, is a key step in the inflammatory response of leukocytes (e.g. basophils, neutrophils, eosinophils, and mast cells). In addition to histamine, these secretory granules contain many proinflammatory mediators including heparin, cytokines, chemokines, and many proteases (Hallgren, 2001). Tryptase, a tetrameric serine proteinase, 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 (He, 2004; He, 1998; Schwartz, 1986). Because it is stored almost exclusively in mast cells, tryptase is a popular indicator of mast cell activation and a target for therapeutic intervention in allergic diseases. The Chemicon Mast Cell Degranulation Assay Kit provides a quick, efficient and sensitive system for evaluation of tryptase activity in cell lysates, supernatants or for inhibitor screening.



Test Principle

Chemicon′s Mast Cell Degranulation Assay Kit provides a simple and convenient means for assaying tryptase activity. The assay is based on spectophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate tosyl-gly-pro-lys-pNA. The free pNA can then be quantified using a spectrophotometer or a microtiter plate reader at 405 nm. A pNA Standard is provided as a comparison control.

Calcium Ionophore A23187 (Calcimycin) is included in the kit for degranulation induction of samples. Calcimycin has been a well-documented inducer of tryptase release from human mast cells (2,3).

A tryptase inhibitor, protamine, is included as a test inhibitor for screening purposes

Quality Level100
ManufacturerSIGMA-ALDRICH
Technique(s)cell based assay: suitable, activity assay: suitable

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