Isakson Lab

Methods

Please feel free to use any of the methods below that are found in the literature.  If you decide the procedures are helpful, we simply ask you cite the papers involved.

Measurement of extracellular ATP release from adherent cells

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ATP release from Red Blood Cells

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Vascular Cell Co-Culture
This method describes a culture model where endothelial cells and vascular smooth muscle cells are cultured on opposing sides of a porous Transwell insert to study myoendothelial junction in vitro. (published: Isakson et al, Circ Res 2005 )

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General Immunocytochemistry
This is our general protocol for performing immunocytochemistry, whether on transverse section of vascular cell co-culture or for monolayers of cells. (published in:  Isakson, J Cell Sci, 2008)

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Paraformaldehyde
This is how we make PFA

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Blocking Solution for Immunocytochemistry
This is our recipe for blocking solution.

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Immunohistochemistry for Electron Microscopy
We use gold beads to identify proteins in EM sections.  (published in:  Isakson, J Cell Sci, 2008)

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Western Blots
We use the LICOR ODYSSEY for development of our western blots to ensure the best possible quantification of protein.  (published in: Johnstone et al, 2012)

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Transfection of Resistance Arteries, Ex Vivo
These are our methods for transfecting intact resistance arteries (in our case, the mouse thoracodorsal arteries) with either siRNA or plasmids, selectivity in smooth muscle cells (published in: Billaud et al, 2011) or endothelial cells (published in: Straub et al, 2012). These arteries are then viable for pressure cannulation and vasoreactivity studies.

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Pressure Myography of Cannulated Arteries

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Membrane Cavolin Fractionation of Cells

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