Mast Cell Tryptase Antibody (Rabbit mAb) [M18M6]

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:100 - 1:250

Application
WB, IP, IHC

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
31 kDa

Positive Control	Human tissue; Human skin; Human colon cancer; Human tonsil
Negative Control

Datasheet & SDS

Biological Description

Specificity
Mast Cell Tryptase Antibody (Rabbit mAb) [M18M6] detects endogenous levels of total Mast Cell Tryptase protein.

Clone
M18M6

Synonym(s)
TPS1, TPS2, TPSB1, TPSAB1, Tryptase alpha/beta-1, Tryptase-1, Tryptase I, Tryptase alpha-1, TPS2, TPSB2, Tryptase beta-2, Tryptase-2, Tryptase II

Background
Mast cell tryptase is the predominant serine protease stored in human mast cell secretory granules, existing not as a free monomeric enzyme but as a heparin-stabilized tetramer assembled from four quasi-equivalent monomers arranged in a flat, frame-like structure; crystallographic analysis shows the four active centers all directed inward toward a narrow central pore roughly 40 by 15 angstroms across, a geometry that inherently restricts access to large macromolecular protein substrates and inhibitors while leaving the enzyme accessible to small peptide substrates, directly explaining tryptase's well-documented resistance to most endogenous proteinaceous protease inhibitors. This tetrameric assembly is not constitutive but requires heparin or related highly sulfated proteoglycans for both activation and stabilization: tryptase tetramers dissociate and lose activity under physiological conditions unless bridged by sufficiently long, negatively charged glycosaminoglycan chains, and this stabilization depends specifically on negative charge density rather than on the particular carbohydrate composition of the polysaccharide, positioning heparin-tryptase complex formation as a structural prerequisite for enzymatic function rather than a mere storage mechanism within the granule. Once released from mast cells during degranulation, tryptase cleaves protease-activated receptor 2 at its N-terminal activating site, generating a tethered ligand that triggers receptor autoactivation, and reconstitution experiments in cells expressing PAR-2 confirm that tryptase-induced receptor activation drives phosphoinositide hydrolysis and elevates cytosolic calcium in cell types that endogenously express PAR-2, including endothelial cells and keratinocytes, with half-maximal signaling occurring at low nanomolar tryptase concentrations. Downstream of PAR-2 activation, tryptase drives fibroblast and airway smooth muscle cell proliferation through a defined signaling cascade requiring the enzyme's active catalytic site, since heat inactivation or active-site-directed protease inhibitors abolish the proliferative response, and this cascade proceeds through a pertussis toxin-sensitive G protein coupled to protein kinase C and tyrosine kinase activity, subsequently inducing cyclooxygenase-2 expression, prostaglandin biosynthesis, and activation of the nuclear receptor PPAR-gamma to drive cell cycle progression.

Background

Mast cell tryptase is the predominant serine protease stored in human mast cell secretory granules, existing not as a free monomeric enzyme but as a heparin-stabilized tetramer assembled from four quasi-equivalent monomers arranged in a flat, frame-like structure; crystallographic analysis shows the four active centers all directed inward toward a narrow central pore roughly 40 by 15 angstroms across, a geometry that inherently restricts access to large macromolecular protein substrates and inhibitors while leaving the enzyme accessible to small peptide substrates, directly explaining tryptase's well-documented resistance to most endogenous proteinaceous protease inhibitors. This tetrameric assembly is not constitutive but requires heparin or related highly sulfated proteoglycans for both activation and stabilization: tryptase tetramers dissociate and lose activity under physiological conditions unless bridged by sufficiently long, negatively charged glycosaminoglycan chains, and this stabilization depends specifically on negative charge density rather than on the particular carbohydrate composition of the polysaccharide, positioning heparin-tryptase complex formation as a structural prerequisite for enzymatic function rather than a mere storage mechanism within the granule. Once released from mast cells during degranulation, tryptase cleaves protease-activated receptor 2 at its N-terminal activating site, generating a tethered ligand that triggers receptor autoactivation, and reconstitution experiments in cells expressing PAR-2 confirm that tryptase-induced receptor activation drives phosphoinositide hydrolysis and elevates cytosolic calcium in cell types that endogenously express PAR-2, including endothelial cells and keratinocytes, with half-maximal signaling occurring at low nanomolar tryptase concentrations. Downstream of PAR-2 activation, tryptase drives fibroblast and airway smooth muscle cell proliferation through a defined signaling cascade requiring the enzyme's active catalytic site, since heat inactivation or active-site-directed protease inhibitors abolish the proliferative response, and this cascade proceeds through a pertussis toxin-sensitive G protein coupled to protein kinase C and tyrosine kinase activity, subsequently inducing cyclooxygenase-2 expression, prostaglandin biosynthesis, and activation of the nuclear receptor PPAR-gamma to drive cell cycle progression.

References
https://pubmed.ncbi.nlm.nih.gov/10500112/ https://pubmed.ncbi.nlm.nih.gov/9020112/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%