The Fc fused LOB7 was used in a 5 μg/mL concentration for detecting vimentin in HUVEC lysates and growth factor reduced matrigel. HUVEC cells harvested from a TC 25 culture flask was lysed using Cell LYTIC M (Sigma Aldrich) according to manufacturer. Growth factor reduced matrigel was thawed and diluted in precooled PBS by factor 10 and 15 μL of this sample was mixed with 5 μL pre-cooled XT sample buffer. The samples were boiled for 5 min. As a positive control, mouse monoclonal anti-vimentin V9 (Sigma Aldrich) was used (1 ug/mL). The primary antibodies were incubated with the blots o/n. For detection of LOB7 and V9, anti-rabbit HRP and anti mouse HRP (DAKO) were incubated with the blots for 1 hr at RT in a 1:1000 dilution.
Mouse monoclonal anti vimentin v9
Manufactured by Merck Group
The Mouse monoclonal anti-vimentin V9 is a laboratory reagent used for the detection and identification of the vimentin protein. Vimentin is an intermediate filament protein found in various cell types. This antibody can be used in a range of immunoassay applications to detect the presence and distribution of vimentin in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cell lytic m, by Merck Group (1 mentions)
Nocodazole, by Merck Group (1 mentions)
Fluorophore conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Blebbistatin, by Merck Group (1 mentions)
Nsc23766, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride, by Merck Group (1 mentions)
Fluorophore conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Y 27632, by Merck Group (1 mentions)
Cytochalasin d, by Merck Group (1 mentions)
2 protocols using mouse monoclonal anti vimentin v9
1
Fc Fused LOB7 Vimentin Detection
2
Immunostaining and Treatment Protocols
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Antibodies included mouse monoclonal anti–vimentin V9 (Sigma-Aldrich, St. Louis, MO), chicken polyclonal anti-vimentin (Novus Biologicals, Littleton, CO), and mouse monoclonal anti–glyceraldehyde-3-phosphate dehydrogenase (Chemicon, Billerica, MA). Fluorophore-conjugated secondary antibodies were used for immunofluorescence (Jackson ImmunoResearch, West Grove, PA). Fluorophore-conjugated phalloidin was used to label polymerized actin (Molecular Probes, Eugene, OR). Cell nuclei were labeled with 4′,6-diamidino-2-phenylindole dihydrochloride (Sigma-Aldrich). For immunoblotting, peroxidase-conjugated secondary antibodies were used (GE Healthcare, Chalfont St Giles, United Kingdom). Cells were treated for 1 h with one or a combination of the following reagents (all from Sigma-Aldrich): blebbistatin (50 μM), cytochalasin D (1 μg/ml), nocodazole (1 μg/ml), Taxol (10 nM; Polioudaki et al., 2009 (link)), Y-27632 (10 μM), NSC23766 (50 μM), and IPA-3 (in 10% dimethylsulfoxide, 10 μM). Controls were performed alongside each treatment and are presented in aggregate.
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