Venous blood was obtained at baseline (0 week visit) and at 8 weeks. Patient plasma was immediately collected and stored at −80°C until use. Parameters measured included: VEGF, tPA, sE- Selectin, bFGF, VCAM-1, and sICAM-1 in patient plasma by ELISA using commercially available Kits (bFGF, VCAM-1, and sICAM-1: RayBiotech Inc, Norcross, GA;. tPA: Abcam, Cambridge, MA; .sE-selectin and VEGF: R&D Systems Inc, Minneapolis, MN; PINP: MyBioSource, San Diego, CA). Proper dilutions were made to ensure that the concentration fell within the standard curve. Samples were all above the detection limit of the assay. Samples were run in duplicate. Anti-human primary antibodies were coated on a 96-well plate before the addition of samples and standards. Sandwich ELISA was completed by adding biotinylated anti-human antibodies, followed by streptavidine-HRP and substrate solution. The optical density of each well was measured using an ELISA plate reader (BioTeK, Winooski, VT).
Vcam 1
Manufactured by RayBiotech
VCAM-1 is a cell surface protein that mediates the adhesion of cells to the vascular endothelium. It plays a role in the inflammatory response and the development of atherosclerosis.
Automatically generated - may contain errors
Lab products found in correlation
Sicam 1, by RayBiotech (2 mentions)
Elisa plate reader, by Agilent Technologies (1 mentions)
Basic fibroblast growth factor (bfgf), by RayBiotech (1 mentions)
Vascular endothelial growth factor (vegf), by RayBiotech (1 mentions)
Svcam 1, by R&D Systems (1 mentions)
Endostatin, by R&D Systems (1 mentions)
Cxcl4, by R&D Systems (1 mentions)
Sflt 1, by R&D Systems (1 mentions)
Se selectin, by R&D Systems (1 mentions)
Basic fibroblast growth factor (bfgf), by R&D Systems (1 mentions)
Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions)
3 protocols using vcam 1
1
Plasma Biomarker Quantification Protocol
2
Cardiac Protein Quantification in Immunized Mice
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Proteins were extracted from hearts of immunized mice and healthy controls. After digestion in 0.1% collagenase for 45 minutes, cells were lysed by ultrasonic pulse echo instrument. cVCAM-1, interleukin-6 (IL-6) and transforming growth factor-β (TGF-β) proteins were quantified using commercially available kits (VCAM-1: RayBiotech, Norcross, GA, Cat# ELM-VCAM1-001; IL-6: Cat#M6000B, TGF-β: Cat#MB100B, R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. For cVCAM-1, each value represents a pooled analysis of two hearts. sVCAM-1 (R&D Systems, Minneapolis, MN, Cat# MVC00, and RayBiotech, Norcross, GA, Cat# ELM-VCAM1-001) was quantified in plasma of immunized mice, sham immunized mice and healthy controls according to the manufacturer’s instructions. For both cardiac and soluble proteins, values represent mean of 2 consecutive measurements.
3
Evaluating Riociguat's Impact on Cardiovascular Biomarkers
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To examine the effect of riociguat on biomarkers, patient plasma was collected at baseline and week 16. Plasma from age- and sex-matched healthy controls was also obtained. The biomarkers were measured by enzyme-linked immunosorbent assays (ELISA) and included total vascular endothelial growth factor (VEGF, R&D Systems), tissue plasminogen activator (tPA, Abcam), soluble E-selectin (sE-selectin, R&D Systems), basic fibroblast growth factor (bFGF, R&D Systems), vascular cell adhesion molecule 1 (VCAM-1, RayBiotech), soluble intracellular adhesion molecule 1 (sICAM-1, RayBiotech), N-terminal propeptide of type I collagen (PINP, MyBioSource), matrix metalloproteinase 12 (MMP12, RayBiotech), CXCL4 (R&D Systems), cyclic guanylyl cyclase (cGMP, Cayman Chemical), endostatin (R&D Systems), and soluble fms-like tyrosine kinase 1 (sFLT1, R&D Systems).
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