Abstract
U87MG cells (human glioblastoma) express tissue factor and shed membrane- derived vesicles enriched in procoagulant activity. Tissue factor antigen has been studied by flow cytometry, immunofluorescent microscopy and Western blotting. Flow cytometric analysis utilized monoclonal antibodies recognizing the tissue factor extracellular domain and the carboxyl-terminal nine amino acids. Studies with intact and permeabilized cells support the location of the carboxyl-terminal domain in the cytoplasm, as previously predicted from the protein sequence. Immunofluorescent microscopy revealed a heterogeneous staining pattern, indicating that tissue factor antigen may be clustered on the cell surface. Intense staining was occasionally observed in cytoplasmic extensions and membrane regions that appeared to be extruding from the cells. Western blot analysis of vesicles shed into the culture medium revealed a principal tissue factor band with mobility marginally slower than that of placental tissue factor. Both extracellular and cytoplasmic epitopes were present in this vesicular tissue factor.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 911-920 |
| Number of pages | 10 |
| Journal | Blood Coagulation and Fibrinolysis |
| Volume | 4 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jan 1 1993 |
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Keywords
- Tissue factor
- U87MG cells
- flow cytometry
- immunohistochemistry
- monoclonal antibodies
ASJC Scopus subject areas
- Hematology
Cite this
Immunofluorescent studies of tissue factor on U87MG cells : Evidence for non-uniform distribution. / Carson, Steven D; Pirruccello, Samuel Jay.
In: Blood Coagulation and Fibrinolysis, Vol. 4, No. 6, 01.01.1993, p. 911-920.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Immunofluorescent studies of tissue factor on U87MG cells
T2 - Evidence for non-uniform distribution
AU - Carson, Steven D
AU - Pirruccello, Samuel Jay
PY - 1993/1/1
Y1 - 1993/1/1
N2 - U87MG cells (human glioblastoma) express tissue factor and shed membrane- derived vesicles enriched in procoagulant activity. Tissue factor antigen has been studied by flow cytometry, immunofluorescent microscopy and Western blotting. Flow cytometric analysis utilized monoclonal antibodies recognizing the tissue factor extracellular domain and the carboxyl-terminal nine amino acids. Studies with intact and permeabilized cells support the location of the carboxyl-terminal domain in the cytoplasm, as previously predicted from the protein sequence. Immunofluorescent microscopy revealed a heterogeneous staining pattern, indicating that tissue factor antigen may be clustered on the cell surface. Intense staining was occasionally observed in cytoplasmic extensions and membrane regions that appeared to be extruding from the cells. Western blot analysis of vesicles shed into the culture medium revealed a principal tissue factor band with mobility marginally slower than that of placental tissue factor. Both extracellular and cytoplasmic epitopes were present in this vesicular tissue factor.
AB - U87MG cells (human glioblastoma) express tissue factor and shed membrane- derived vesicles enriched in procoagulant activity. Tissue factor antigen has been studied by flow cytometry, immunofluorescent microscopy and Western blotting. Flow cytometric analysis utilized monoclonal antibodies recognizing the tissue factor extracellular domain and the carboxyl-terminal nine amino acids. Studies with intact and permeabilized cells support the location of the carboxyl-terminal domain in the cytoplasm, as previously predicted from the protein sequence. Immunofluorescent microscopy revealed a heterogeneous staining pattern, indicating that tissue factor antigen may be clustered on the cell surface. Intense staining was occasionally observed in cytoplasmic extensions and membrane regions that appeared to be extruding from the cells. Western blot analysis of vesicles shed into the culture medium revealed a principal tissue factor band with mobility marginally slower than that of placental tissue factor. Both extracellular and cytoplasmic epitopes were present in this vesicular tissue factor.
KW - Tissue factor
KW - U87MG cells
KW - flow cytometry
KW - immunohistochemistry
KW - monoclonal antibodies
UR - http://www.scopus.com/inward/record.url?scp=0027723682&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027723682&partnerID=8YFLogxK
U2 - 10.1097/00001721-199312000-00008
DO - 10.1097/00001721-199312000-00008
M3 - Article
VL - 4
SP - 911
EP - 920
JO - Blood Coagulation and Fibrinolysis
JF - Blood Coagulation and Fibrinolysis
SN - 0957-5235
IS - 6
ER -