Saphenous vein-derived ECs and VSMCs were separately cultured on 6-well plates and treated with TGFβ1 (0, 1, and 10 ng/ml) for 24 h. The resulting conditioned medium was harvested and used to determine the relative levels of AGFs by ELISA (VEGF-C, Ang-1, TGF-β1; R&D Systems Ltd) or Quantibody multiplex array (Angiogenin, Ang-2, EGF, bFGF, HB-EGF, HGF, Leptin, PDGF-BB, PlGF, VEGF-A; Raybiotech, Norcross, United States) according to the manufacturers’ instructions. Each experiment was performed on three separate occasions.
Pdgf bb
Manufactured by RayBiotech
Sourced in United States
PDGF-BB is a recombinant human platelet-derived growth factor BB homodimer. It is a signaling protein involved in cell growth and division.
Automatically generated - may contain errors
Lab products found in correlation
Basic fibroblast growth factor (bfgf), by RayBiotech (2 mentions)
Leptin, by RayBiotech (1 mentions)
Vegf a, by RayBiotech (1 mentions)
Hepatocyte growth factor (hgf), by RayBiotech (1 mentions)
Hb egf, by RayBiotech (1 mentions)
Ang 2, by RayBiotech (1 mentions)
Angiogenin, by RayBiotech (1 mentions)
Epidermal growth factor (egf), by RayBiotech (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Vegf c, by R&D Systems (1 mentions)
Ang 1, by R&D Systems (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Advia 2120, by Bayer (1 mentions)
Vegf elisa kit, by R&D Systems (1 mentions)
Timp 4, by RayBiotech (1 mentions)
Tgf α, by RayBiotech (1 mentions)
Timp 2, by RayBiotech (1 mentions)
Timp 1, by RayBiotech (1 mentions)
Tgfβ1, by RayBiotech (1 mentions)
5 protocols using pdgf bb
1
Profiling Angiogenic Factors in EC/VSMC
2
VSMC Viability Assay via MTT
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Cell viability was determined by MTT assay. Human umbilical vein VSMCs were seeded in 96-well plates at a density of 5×104 cells/well. After PDGF-BB (RayBiotech, USA) incubation, the cells were washed twice with PBS, followed by incubation with 25 μl of MTT (Sigma, USA) at 37C for 4 h in the dark. Subsequently, 150 ml of DMSO (Sigma, USA) was added to dissolve MTT crystals. The absorbance was measured in a microplate reader at 490 nm (Bio-Rad 680, USA).
3
Platelet-Rich Plasma Biomarker Analysis
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The white blood cell, platelet, lymphocyte, neutrophil, monocyte, and red blood cell counts of the PRP and plasma were measured using a blood analyzer (Advia 2120, Bayer); as described in our previous study22 (link), several growth factors were detected using an enzyme-linked immunosorbent assay (ELISA). The standard curve was obtained for each experiment. Platelet-derived growth factor (PDGF) AA, PDGF BB (RayBiotech Co., Georgia, USA), epidermal growth factor (EGF), and vascular epidermal growth factor (VEGF; Koma Co., Republic of Korea) levels were quantitatively measured using ELISA kits, with reference range was from 0 to 3000 (PDGF AA), from 0 to 2000 (PDGF BB), from 0 to 1000 (EGF) and from 0 to 1000 (VEGF) pg/ml, respectively. All measurements were conducted in triplicate.
4
Quantifying Wound Healing Factors
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Specific components relevant to wound healing were identified and concentrations of these components were measured using enzyme-linked immunosorbent assay (ELISA) in each sample type. PDGF-BB, PDGF-AA, TIMP-1, TIMP-2, TIMP-4, TGF-β1, TGF-α, bFGF, and EGF ELISA kits were obtained from RayBiotech, Inc (Norcross, GA). Hyaluronan (Hyaluronic acid [HA] ) and VEGF ELISA kits were obtained from R&D Systems (Bio-Techne Corporation, Minneapolis, MN). Lactoferrin ELISA kits were obtained from AssayPro, LLC (St. Charles, MO). Assays were performed according to each kit manufacturer's instructions. Component concentrations in the IL are reported. To compare component concentrations, dCHPM component concentrations were normalized to the component concentrations of fresh, unprocessed PM.
5
Stem Cell Culture and Growth Factor Analysis
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Dispase II was obtained from Roche Diagnostics Corporation (Indianapolis, IN). Phosphate-buffered saline (PBS), trypsin-EDTA, F12, Dulbecco’s modified Eagle’s medium (DMEM), amphotericin B and FBS were purchased from Gibco (Rockville, MD). Enzyme-linked immunosorbent assay (ELISA) kit for human EGF kit was purchased from eBioscience (San Diego, CA). ELISA kits for TGF-β1, PDGF-AB and PDGF-BB were acquired from RayBiotech, Inc. (Norcross, GA), R&D Systems (Minneapolis, MN) and PeproTech (Rocky Hill, NJ), respectively. All other reagents were obtained from Sigma-Aldrich (St. Louis, MO).
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