2012 Promega catalogue

Cell Signaling
Tryptase, Human, Recombinant, β
Product Size Cat.# Price ($)
rhSkin β Tryptase 100 µg G7061 801.00
rhLung β Tryptase 100 µg G5631 801.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: Tryptase is the predominant protein in mast cell granules and
cleaves proteins at arginine and lysine residues. Tryptase is stored and released
from mast cell granules upon activation. Mast cells are found in many tissues
but are present in greater numbers along epithelial linings of the body, such as
the skin, respiratory tract and gastrointestinal tract, as well as the perivascular
tissue surrounding blood vessels. They are involved in a variety of physiological
and pathophysiological states, including immediate hypersensitivity, delayedtype
hypersensitivity, cell growth regulation, defense against neoplasia, and
pain and itch sensation. They have also been implicated in chronic
inflammatory states and are involved in neuroimmune interactions. The
availability of recombinant human tryptase will aid in research directed toward
a more complete understanding of the biological role(s) of tryptase and mast
cells and the identification of tryptase in vivo targets.
Skin β I Tryptase, Human, Recombinant (rhSkin β Tryptase) and Lung β II
Tryptase, Human, Recombinant (rhLung β Tryptase) are neutral serine
proteases. The human β tryptase enzymes have been cloned and stably
expressed in Pichia pastoris as fully active tetrameric enzymes and purified by
affinity chromatography. The two enzymes differ in buffer formulation, enzyme
concentration and glycosylation pattern. rhLung Tryptase is provided at a much
higher concentration (2mg/ml) in minimal buffer without heparin for
chromatographic studies and with glycosylation more closely resembling
cadaveric enzyme as demonstrated by glycosidase digestion followed by
Western analysis of the two recombinant enzymes and native lung tryptase.
Specific Activity: Measured as the rate of hydrolysis of 0.4mM Nα
CBZ-L-Lysine Thiobenzyl Ester as substrate coupled with Ellman’s Reagent
(5,5´-Dithio-bis(5-Nitrobenzoic Acid)) in a final volume of 1ml, incubating for 1
minute at 25°C, and monitoring the absorbance change at 410nm. One unit is
defined as 1.0 absorbance unit change per minute.
• rhSkin β Tryptase: >1,000 units/mg protein.
• rhLung β Tryptase: >1,200 units/mg protein.
Concentration:
• rhSkin β Tryptase: 200μg/ml.
• rhLung β Tryptase: 2mg/ml.
Features:
• High Specific Activity: Specific activity is consistently 130–150% higher
than native lung tryptase.
• Consistent: Recombinant protein expression results in uniform enzyme
from batch to batch.
• Safe: Void of human pathogens associated with native cadaveric tryptase.
• Pure: Skin β and Lung β Tryptase are free of other contaminating proteases,
providing more active enzyme per milligram of protein and eliminating
extraneous protein interactions observed with native tryptase.
Storage Conditions: Store at –20°C.
For complete and up-to-date product information visit: www.promega.com/catalog
undigested digested
MW
Native
rhSkin
rhLung
Native
rhSkin
rhLung
MW
—34kDa
—29kDa
Glycosidase digestion of human β tryptase with PNGase F yields
single tryptase core protein. Western blot with Anti-Human Tryptase mAb
(clone AA5, Cat.# G3361).
ADME Assays
For additional information see page 20.
Apoptosis
For additional information see page 25.
Cell Viability Assays
For additional information see page 34.
Cytotoxicity Assays
For additional information see page 38.
Glutathione Measurement
For additional information see page 44.
Histone Deacetylase Assays
For additional information see page 52.
Nuclear Receptor Pathway Tools
For additional information see page 271.
Protein Kinases and Kinase Assays
For additional information see page 64.
3521TA08_1A
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