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Rabbit anti ve cadherin

Manufactured by Santa Cruz Biotechnology
Sourced in United States

Rabbit anti-VE-cadherin is a primary antibody that recognizes the vascular endothelial (VE)-cadherin protein. VE-cadherin is an adhesion molecule that plays a crucial role in the formation and maintenance of endothelial cell-cell junctions. This antibody can be used to detect and study the expression and localization of VE-cadherin in various experimental settings.

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5 protocols using rabbit anti ve cadherin

1

Western Blot Protein Analysis Protocol

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The protein content of cell extracts or tissue prepared in radioimmunoprecipitation assay lysis buffer was determined using the Bradford protein assay kit (Bio-Rad). About 30 μg of proteins were separated by SDS-polyacrylamide gel electrophoresis. After transferring to nitrocellulose membranes, immunoblots were probed separately with various primary antibodies. Subsequently, the immunoblots were blocked with 5% skimmed milk in Tris-buffered saline solution. Fluorescently labeled or horseradish peroxidase–conjugated secondary antibodies were detected by the Odyssey Infrared Imaging System (LI-COR Biosciences). Primary antibodies were used as following: rabbit anti-JMJD3 (Millipore; catalog no.: 07-1533), rabbit anti-H3K27me3/me2/me1 (Abclonal; catalog nos.: A2363, A2362, and A2361), rabbit anti-UTX (GeneTex; catalog no.: GTX12146), rabbit anti-eNOS (Cell Signaling Technology; catalog no.: 32027), rabbit anti-VE-cadherin (Santa Cruz; catalog no.: sc-28644), mouse anti-α-SMA (Sigma; catalog no.: A5228), goat anti-Hes1 (Santa Cruz; catalog no.: sc-13844), rabbit anti-PCNA (Santa Cruz; catalog no.: sc-7907), mouse anti-β-actin (GeneTex; catalog no.: GTX629630), and mouse anti-GAPDH (Santa Cruz; catalog no.: sc-32233).
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2

Endothelial Cell Permeability Assay

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The following were used: LPS from Escherichia coli O111:B4, O55:B5, thrombin, and TNF-α (Sigma Aldrich, USA); X-tremeGENE siRNA Transfection Reagent and X-tremeGENE HP DNA Transfection Reagent (Roche, Switzerland); EntransterTM-in vivo transfection reagent (Engreen Biosystem, China); rhodamine-phalloidin (Invitrogen, USA); Rab5a activation assay kit (NewEast Biosciences, USA); FITC-dextran and chloroquine (Santa Cruz, USA); rabbit anti-VE-cadherin, anti-Podxl, goat anti-VE-cadherin, and mouse anti-CD31 (Santa Cruz, USA); rabbit anti-Rab5 (ABCam, USA); rabbit anti-β-actin (Cell Signaling Technology, USA); rabbit anti-F-actin and anti-GAPDH (Biosynthesis Biotechnology, Beijing, China); Alexa Fluor 488- or Alexa Fluor 594-labeled and HRP-coupled goat anti-mouse and anti-rabbit IgG and Dylight 649-donkey anti-goat IgG (ZSGB-Bio, Beijing, China).
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3

Immunostaining of Cellular Markers

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Immunostaining was performed as previously described in detail (Jumabay et al., 2012 (link)). Briefly, cells grown in chamber slides were fixed in 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, blocked with 10% goat serum and 1% BSA in PBS, and incubated over night at 4°C with the appropriate primary antibodies or non-specific IgG control antibodies, diluted 1:200 in 1% BSA in PBS. The next day, cells were incubated with secondary AF-488-conjugated (green fluorescence) or AF-594-conjugated (red fluorescence) goat anti-mouse or anti-rabbit secondary antibodies (Molecular Probes). The cells were washed with PBS, the nuclei stained with 4',6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich), and visualized by fluorescence microscopy. The non-specific IgG control antibodies showed no staining and are not included in the figures.
We used the following antibodies for immunostaining: hamster anti-CD31, rabbit anti-vone Willebrand Factor (vWF) (both from Dako), goat anti-BMP4, goat anti-MGP, rabbit anti-VEGF, rabbit anti-VE-Cadherin (all from Santa Cruz Biotechnology), mouse anti-Perilipin (Cell Signaling Technology).
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4

Vascular Smooth Muscle Cell Characterization

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Collagenase type II, Elastase, and Exisulind were obtained from Sigma-Aldrich co. (St Louis, MO, USA). PDGF-BB was purchased from R&D systems (Minneapolis, MN, USA). Goat anti-PKG I, rabbit anti-VE-cadherin, mouse anti-osteoponin, rabbit anti-CD31, and goat anti-actin antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-VASP (Ser239), rabbit anti-t-Akt, and mouse anti-p-Akt (Ser473) were purchased from Cell Signaling (Berkely, MA, USA). Mouse anti-calponin antibody was obtained from Sigma-Aldrich co. (St Louis, MO, USA). Mouse anti-α-SMA antibody was purchased from Abcam (Cambridge, MA, USA). The secondary antibodies to each primary antibody were as follows. Anti-mouse IgG HRP conjugated and anti-rabbit IgG HRP conjugated antibodies were obtained from Promega (Madison, WI, USA). Anti-goat IgG HRP conjugated antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-mouse IgG Alexa flour 488 and anti-rabbit IgG Alexa flour 594 secondary antibodies were purchased from Invitrogen (Carlsbad, CA, USA). Alzet osmotic pump was purchased from Durect corporation (Cupertino, CA, USA).
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5

Endothelial Cell Permeability Assay

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The following were used: LPS from Escherichia coli O111:B4, O55:B5, thrombin, and TNF-α (Sigma Aldrich, USA); X-tremeGENE siRNA Transfection Reagent and X-tremeGENE HP DNA Transfection Reagent (Roche, Switzerland); EntransterTM-in vivo transfection reagent (Engreen Biosystem, China); rhodamine-phalloidin (Invitrogen, USA); Rab5a activation assay kit (NewEast Biosciences, USA); FITC-dextran and chloroquine (Santa Cruz, USA); rabbit anti-VE-cadherin, anti-Podxl, goat anti-VE-cadherin, and mouse anti-CD31 (Santa Cruz, USA); rabbit anti-Rab5 (ABCam, USA); rabbit anti-β-actin (Cell Signaling Technology, USA); rabbit anti-F-actin and anti-GAPDH (Biosynthesis Biotechnology, Beijing, China); Alexa Fluor 488- or Alexa Fluor 594-labeled and HRP-coupled goat anti-mouse and anti-rabbit IgG and Dylight 649-donkey anti-goat IgG (ZSGB-Bio, Beijing, China).
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