Rabbit polyclonal anti‐α‐SMA was acquired from Abcam. Rabbit polyclonal anti‐GAPDH, anti‐von Willebrand Factor (vWF), and anti‐fibroblast‐specific protein 1 (FSP1) were acquired from Santa Cruz Biotechnology. Rabbit Polyclonal Endothelin‐1 was purchased from Fisher Scientific. Rabbit polyclonal anti‐BMPRII and anti‐P‐SMAD1/5/8 were purchased from Cell Signaling Technology. Mouse monoclonal anti‐eNOS, anti‐β‐actin and rabbit polyclonal anti‐Cav‐1 were purchased from BD PharMingen. Alexa‐Fluor 488 and 555‐conjugated goat and donkey antimouse or antirabbit IgG were purchased from Life Technologies. Antimouse and antirabbit HRP‐conjugated IgG were purchased from Cell Signaling Technology or Kierkegaard and Perry Laboratories (Supporting Information 1 ). RIPA buffer, protease, and phosphatase inhibitor cocktail, collagenase type I, paraformaldehyde, heparin, and sucrose were purchased from SIGMA Chemical, Co. Mounting media containing DAPI (VectaShield) was obtained from Vector. Stock solutions were prepared in 100% dimethylsulfoxide (DMSO), buffered physiological solution or sterile phosphate‐buffered saline (PBS) and diluted daily in sterile PBS or DMEM. The highest final concentration of the solvent was 0.1% (v/v) and did not affect the experiments. PCR primers were purchased from Integrated DNA Technologies, Inc.
Anti von willebrand factor
Manufactured by Santa Cruz Biotechnology
Anti‐von Willebrand Factor (vWF) is a protein that plays a crucial role in blood clotting. It is responsible for facilitating the adhesion of platelets to the site of vascular injury, which is an essential step in the process of hemostasis.
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Lab products found in correlation
Protease and phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Heparin, by Merck Group (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Anti bmprii, by Cell Signaling Technology (1 mentions)
Anti p smad1 5 8, by Cell Signaling Technology (1 mentions)
Anti cav1, by BD (1 mentions)
Antimouse and antirabbit hrp conjugated igg, by Cell Signaling Technology (1 mentions)
Anti α sma, by Abcam (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Anti β actin, by BD (1 mentions)
Magnetic column, by Miltenyi Biotec (1 mentions)
Fibronectin coated chamber slides, by BD (1 mentions)
Streptavidin conjugated magnetic beads, by Miltenyi Biotec (1 mentions)
Anti endothelial nitric oxide synthase, by BD (1 mentions)
Dii ac ldl, by Thermo Fisher Scientific (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
2 protocols using anti von willebrand factor
1
Antibody and Reagent Sources for Cell Analysis
2
Magnetic Separation of CD45+ Cells
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Cells derived from high density culture at day 15 were stained with a biotinylated anti-rat-CD45 monoclonal antibody (BD Biosciences) for 30 min on ice, washed, and then incubated with streptavidin-conjugated magnetic beads (Miltenyi Biotec, Teterow, Germany) for 20 min. Cells were then separated into CD45+ and CD45− cells using magnetic columns (Miltenyi Biotec) and plated onto fibronectin-coated chamber slides (BD Biosciences) in EGM-2 medium supplemented with 50 ng/ml vascular endothelial cell growth factor (VEGF, Lonza, Walkersville, MD, USA). After 14 days of induction, cells were incubated with DiI-ac-LDL (Invitrogen) as previously described, or fixed and stained with anti-von Willebrand factor (vWF) (Santa Cruz Biotechnology) or anti-endothelial nitric oxide synthase (eNOs) (BD Biosciences) antibodies and visualized with an Alexa Fluor 488-conjugated secondary antibody. Three representative fields were recorded and the positive cells were calculated by Image-Pro Plus software (Media Cybernetics, Rockville, MD, USA).
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