Katherine R. Yates, Jessica Welsh, Hussein H.
Reversible Coagulation in Living Tissue: VIII
Pancreatic cancer cell and microparticle procoagulant surface characterization. Low-carb tissue factor?. Journal of Thrombosis and Haemostasis , 9 8 , Glycosylation of tissue factor is not essential for its transport or functions. Jolanta Krudysz-Amblo, Mark E. Jennings, Dwight E. Matthews, Kenneth G. Mann, Saulius Butenas. Differences in the fractional abundances of carbohydrates of natural and recombinant human tissue factor. The missing link between atherosclerosis, inflammation and thrombosis: is it tissue factor?.
Expert Review of Cardiovascular Therapy , 9 4 , Hussein Echrish, Leigh A. Madden, John Greenman, Anthony Maraveyas. Cancers , 3 1 , Posttranslational modifications and activity of natural and recombinant tissue factor. Thrombosis Research , , SS Narcis I. Popescu, Cristina Lupu, Florea Lupu. Jennings, Kenneth G. Journal of Biological Chemistry , 5 , Tissue Factor in Coagulation. Arteriosclerosis, Thrombosis, and Vascular Biology , 29 12 , Hollingshead, J.
Gerard Wall. Applied Biochemistry and Biotechnology , 1 , Tissue factor activity and function in blood coagulation. Elena M. Egorina, Mikhail A. Regulation of tissue factor procoagulant activity by post-translational modifications. Thrombosis Research , 6 , Brummel-Ziedins, Kenneth G. Tissue factor in thrombosis and hemorrhage. Surgery , 4 , S2-S Chao Li, Lucy M. Colman, Mary E. Collier, Charlotte E. Dyer, John Greenman, Camille Ettelaie. Tumour-expressed tissue factor inhibits cellular cytotoxicity.
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Cancer Immunology, Immunotherapy , 55 11 , Karl-Erik Eilertsen, Bjarne?? Tissue factor. Lopes-Bezerra, S. Endothelial cells, tissue factor and infectious diseases.
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Brazilian Journal of Medical and Biological Research , 36 8 , Expression of recombinant rabbit tissue factor in Pichia pastoris, and its application in a prothrombin time reagent. Protein Expression and Purification , 26 3 , Paolo Golino. The inhibitors of the tissue factor:factor VII pathway. Thrombosis Research , 3 , VV Esther Bachli, MD. Historical review. British Journal of Haematology , 2 , John H. Tissue Factor pathway. David M.
Tissue Factor and Biotechnology. Thrombosis Research , 90 1 , Masaki Iino, Donald C. Foster, Walter Kisiel.
Archives of Biochemistry and Biophysics , 2 , Radha Gudepu Krishna, Finn Wold. Posttranslational Modifications. Annet Hammacher, Gavin E. Reid, Robert L. Moritz, Richard J. Identification and characterization of recombinant murine interleukin-6 with a C-terminal pentapeptide extension using capillary reversed phase HPLC-MS and Edman degradation. Biomedical Chromatography , 11 6 , Magnus Jansson, Dan Hall? Journal of Biological Chemistry , 13 , Expression, purification, and characterization of Escherichia coli-derived recombinant human melanoma growth stimulating activity.
Christine D. McCallum, Raymond C. Hapak, Pierre F. Neuenschwander, James H. Morrissey, Arthur E. Journal of Biological Chemistry , 45 , Qiuling Huang, Pierre F. Neuenschwander, Alireza R. Rezaie, James H. Journal of Biological Chemistry , 36 , Gregory A. Kreutzer, Phillipe de Moerloose, Andrew W. Boyd, Irene Stanley, Anthony W. Burgess, Frederick R. Tissue factor expression in human leukemic cells. Leukemia Research , 20 1 , Richard J.
Dorow, Gavin E. Moritz, Guo-Fen Tu. Identification of truncated E. The final step of the purification involves size exclusion chromatography using a Sephacryl S column. Protein concentrations were measured using the bicinchoninic acid assay and UV-Vis method A with A correction.
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Neutral red is a vital dye that is taken up by live cells, the absence of red denotes a plaque. Plaques are counted and multiplied by the dilution factor to obtain the plaque forming units pfu , a unit of virus infectivity. Protein staining was achieved by using Coomassie R protein stain.
B Improvements to previous protocols included heparin column prior to metal-ion affinity and size exclusion post-metal-ion affinity chromatography and resulted in a cleaner preparation. Molecular weight M. The spacer sequence was included to help orient the protein when bound to an appropriate membrane surface. Briefly, E. Transformants were selected with kanamycin and were grown in a 16x mm test tube containing 1.
Following IPTG induction and overnight incubation at room temperature rpm shaking , the media was collected and centrifuged to remove cells and cell debris. Osmotic shock was performed on the pelleted cells to release sTF stuck in the periplasmic space of the bacteria . Following dialysis of the precipitate, the protein solution was batch-treated with Q Sepharose. The unbound solution contained sTF-His and was applied to an immobilized nickel ion affinity resin. Anti-TF , ab was used as the primary antibody with goat anti-rabbit HRP , as the secondary antibody.
This method relies on antibody recognition of an antigen with visualization of these antigens via dense colloidal gold. Colloidal gold appears as dark dots by EM and can be generated in different sizes. Conjugation of gold colloidal to antibodies either primary or secondary can be easily achieved through passive adsorption or covalent chemistry.
For my cell-cultured virus studies, gold visualization was performed using commercial secondary antibodies conjugated to gold particles of varying diameters 6 nm, 10 nm and 15 nm. For patient-derived virus studies, in-house conjugation of primary antibodies was performed. This was important due to the lack of host selection of antibodies against virus proteins which would complicate simultaneous labeling of antigens on the virus surface.
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