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Protein assay materials were purchased from Bio-Rad Laboratories (Hercules, CA).
To evaluate the effective of HSPG binding peptide in endothelial cell migration, we used rhVEGF165 to induce HUVECs migration in a transwell assay (Materials and Methods).
Protein assay materials and SYBR Green Supermix were purchased from Bio-Rad Laboratories (Hercules, CA), and Geneticin was from Life Technologies Corporation (Carlsbad, CA).
To evaluate the ability of apoEdp in blocking VEGF-induced HUVEC migration and invasion, we used a Boyden chamber transwell assay (Materials and Methods).
The binding and uptake of TfR was kinetically analyzed over 2 hrs with saturating levels of A568Tf (Figure 3A) in an approach to steady-state assay (Materials and Methods; [32]).
All of the assay materials and reagents were provided by the Beckman Coulter Ins.
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Muscle biopsy is the most common diagnostic tool for assay material.
Mass-spec and RT-PCR quantifications will be robust by the consistency of the assay material.
Blood would most likely be the assay material of choice and this in itself may present limitations if responses are localized to deeper tissues.
To compare Myogel with Matrigel® (BD Matrigel Matrix, BD Biosciences, Cat. Number 354234) as an invasion assay material, experiments were carried out according to BD Biosciences instructions to coat the filter membranes with Matrigel®.
As FGF2 itself is released upon explantation of cartilage [ 5], this process may affect the balance of FGFRs in the resulting assay material, perhaps leading to skewing of the natural balance of receptors and tending towards a more catabolic pheno-type in cultured cells/explants in vitro.
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