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Sedlmayr, P; Grosshaupt, B; Muntean, W.
Flow cytometric detection of intracellular platelet antigens.
Cytometry. 1996; 23(4):284-289 Doi: 10.1002/(SICI)1097-0320(19960401)23:4<284::AID-CYTO4>3.0.CO;2-H [OPEN ACCESS]
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Leading authors Med Uni Graz
Sedlmayr Peter
Co-authors Med Uni Graz
Muntean Eugen
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Abstract:
Exocytosis following platelet activation leads to translocation of CD62P (P-selectin), CD63, and thrombospondin from cytoplasmic granules to the cell surface membrane, where these molecules, serving as activation markers, can be detected by flow cytometry. We report the detectability of these molecules preformed--prior to platelet activation--inside the cytoplasm of resting platelets. Two different methods are described, using either methanol or the Fix&Perm kit for cell membrane permeabilization. In addition, interleukin(IL-)1 alpha is shown to be present in platelet cytoplasm after methanol treatment but not after permeabilization using Fix&Perm. Whenever cell surface positivity for a specific marker coincides with intracellular presence, blocking of the surface membrane sites prior to membrane permeabilization is required in order to obtain fluorescence intensity attributable to cytoplasmic staining only. Our data demonstrate the feasibility of the methods described for the detection of intracellular platelet molecules.
Find related publications in this database (using NLM MeSH Indexing)
Adult -
Antigens, CD - analysis
Blood Platelets - immunology
Flow Cytometry - immunology
Humans - immunology
Interleukin-1 - analysis
Membrane Glycoproteins - analysis
P-Selectin - analysis
Platelet Membrane Glycoproteins - analysis
Thrombospondins - analysis

Find related publications in this database (Keywords)
Flow Cytometry Methods
Cell Membrane Permeabilization
Intracellular
Cd62P
P-Selectin
Cd63
Thrombospondin
Interleukin-1-Alpha
Cytokine
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