rabbit anti trpc 1  (Alomone Labs)

Journal: Arthritis Research & Therapy

Article Title: Proangiogenic effects of soluble α-Klotho on systemic sclerosis dermal microvascular endothelial cells

doi: 10.1186/s13075-017-1233-0

Figure Lengend Snippet: Schematic representation of the molecular complex formed by vascular endothelial growth factor ( VEGF )-bound VEGF receptor-2 ( VEGFR-2 ), transient receptor potential canonical-1 ( TRPC-1 ) and soluble α-Klotho ( sKl ) on the surface of endothelial cells ( ECs ). This molecular complex regulates the influx of Ca 2+ and the permeability of the plasma membrane contributing to the maintenance of EC integrity

Article Snippet: The membranes were blocked with blocking solution included in the Western Breeze Chromogenic Western Blot Immunodetection Kit (Invitrogen) for 40 minutes at room temperature on a rotary shaker and subsequently incubated for 1 hour at room temperature with rabbit monoclonal anti-human α-Klotho (1:500 dilution; catalog number ab181373; Abcam), mouse monoclonal anti-human VEGF 165 b (1:500 dilution; catalog number ab14994; Abcam), rabbit polyclonal anti-human VEGFR-2 (1:100 dilution; catalog number ab45010; Abcam), rabbit monoclonal anti-human TRPC-1 (1:1000 dilution; catalog number ab51255; Abcam), rabbit monoclonal anti-human Ki67 (1:1000 dilution; catalog number ab92742; Abcam), rabbit polyclonal anti-human caspase-3 (1:1000 dilution; catalog number 9662; Cell Signaling Technology, Danvers, MA, USA), and rabbit polyclonal anti-human α-tubulin (1:1000 dilution; catalog number ab18251; Abcam) antibodies, assuming α-tubulin as control invariant protein.

Techniques: Permeability

Journal: Arthritis Research & Therapy

Article Title: Proangiogenic effects of soluble α-Klotho on systemic sclerosis dermal microvascular endothelial cells

doi: 10.1186/s13075-017-1233-0

Figure Lengend Snippet: Expression of α-Klotho, vascular endothelial growth factor ( VEGF ) 165 b, VEGF receptor-2 ( VEGFR-2 ) and transient receptor potential canonical-1 ( TRPC-1 ) in dermal microvascular endothelial cells from healthy controls ( H-MVECs ) and patients with systemic sclerosis (SSc) ( SSc-MVECs ). a , b Representative microphotographs of immunocytochemical staining for α-Klotho in H-MVECs ( a ) and SSc-MVECs ( b ) detected with biotinylated secondary antibodies and streptavidin peroxidase. c - f Protein lysates from H-MVECs and SSc-MVECs were subjected to western blotting analysis using anti-α-Klotho ( c ), anti-VEGF 165 b ( d ), anti-VEGFR-2 ( e ) and anti-TRPC-1 ( f ) antibodies. Representative immunoblots are shown. The densitometric analysis of the bands normalized to α-tubulin is reported in the histograms. Data are mean ± SD of optical density ( OD ) in arbitrary units. Student’s t test was used for statistical analysis; p values are indicated in each panel. Results are representative of three independent experiments performed with each one of the five H-MVEC and five SSc-MVEC lines

Article Snippet: The membranes were blocked with blocking solution included in the Western Breeze Chromogenic Western Blot Immunodetection Kit (Invitrogen) for 40 minutes at room temperature on a rotary shaker and subsequently incubated for 1 hour at room temperature with rabbit monoclonal anti-human α-Klotho (1:500 dilution; catalog number ab181373; Abcam), mouse monoclonal anti-human VEGF 165 b (1:500 dilution; catalog number ab14994; Abcam), rabbit polyclonal anti-human VEGFR-2 (1:100 dilution; catalog number ab45010; Abcam), rabbit monoclonal anti-human TRPC-1 (1:1000 dilution; catalog number ab51255; Abcam), rabbit monoclonal anti-human Ki67 (1:1000 dilution; catalog number ab92742; Abcam), rabbit polyclonal anti-human caspase-3 (1:1000 dilution; catalog number 9662; Cell Signaling Technology, Danvers, MA, USA), and rabbit polyclonal anti-human α-tubulin (1:1000 dilution; catalog number ab18251; Abcam) antibodies, assuming α-tubulin as control invariant protein.

Techniques: Expressing, Staining, Western Blot

Journal: Arthritis Research & Therapy

Article Title: Proangiogenic effects of soluble α-Klotho on systemic sclerosis dermal microvascular endothelial cells

doi: 10.1186/s13075-017-1233-0

Figure Lengend Snippet: Expression of vascular endothelial growth factor ( VEGF ) 165 b and transient receptor potential canonical-1 ( TRPC-1 ) in dermal microvascular endothelial cells from healthy controls ( H-MVECs ) and systemic sclerosis (SSc) patients ( SSc-MVECs ). a , b Western blotting of total protein extracts from H-MVECs and SSc-MVECs at basal condition and treated with recombinant human soluble α-Klotho ( sKl ) protein for 24 and 48 hours assayed with anti-VEGF 165 b ( a ) and anti-TRPC-1 ( b ) antibodies. Representative immunoblots are shown. The densitometric analysis of the bands normalized to α-tubulin is reported in the histograms. Data are mean ± SD of optical density ( OD ) in arbitrary units. Student’s t test was used for statistical analysis; * p < 0.05 versus basal H-MVECs, ° p < 0.05 versus basal SSc-MVECs. Results are representative of three independent experiments performed with each one of the five H-MVEC and five SSc-MVEC lines

Article Snippet: The membranes were blocked with blocking solution included in the Western Breeze Chromogenic Western Blot Immunodetection Kit (Invitrogen) for 40 minutes at room temperature on a rotary shaker and subsequently incubated for 1 hour at room temperature with rabbit monoclonal anti-human α-Klotho (1:500 dilution; catalog number ab181373; Abcam), mouse monoclonal anti-human VEGF 165 b (1:500 dilution; catalog number ab14994; Abcam), rabbit polyclonal anti-human VEGFR-2 (1:100 dilution; catalog number ab45010; Abcam), rabbit monoclonal anti-human TRPC-1 (1:1000 dilution; catalog number ab51255; Abcam), rabbit monoclonal anti-human Ki67 (1:1000 dilution; catalog number ab92742; Abcam), rabbit polyclonal anti-human caspase-3 (1:1000 dilution; catalog number 9662; Cell Signaling Technology, Danvers, MA, USA), and rabbit polyclonal anti-human α-tubulin (1:1000 dilution; catalog number ab18251; Abcam) antibodies, assuming α-tubulin as control invariant protein.

Techniques: Expressing, Western Blot, Recombinant