The Only Method for Direct Quantification of Vascular Nitric Oxide (EDRF)

Samples

• Mouse
  1.5 cm e. g. aorta cut in 5 pieces
• Rat
  1.0 cm e. g. aorta cut in 3 pieces
• Rabbit
  0.5 cm e. g. vena cava in a piece
• In general
  50-100 mm2 intimal surface of the
  vessel, about 1 - 2 x 105
  endothelial cells

Basal (orange) and calcium ionophor stimulated (blue) nitric oxide (NO) generation of rat aorta

Detailed information about this method

• Advanced spin Trapping of vascular nitric Oxide using colloid iron diethyldithiocarbamate.
  Kleschyov, A. L., Münzel, T., Methods Enzymol. 359, 42-51 (2002).
• Spin trapping of vascular nitric oxide using colloid Fe(II)-diethydithiocarbamate.
  Kleschyov, A. L., Mollnau, H., Oelze, M., Meinertz, T., Huang, Y., Harrison, D.G., Münzel, T., Biochem. Biophys. Res. Commun. 275, 672-677 (2000).

Spin Trap

• 400 µM colloid Fe(II)-Diethyldithiocarbamate, [Fe(II)(DETC)₂]
• Applied end concentration
  50 - 250 µM [Fe(II)(DETC)₂]
• Time for sample incubation
  15 - 60 minutes at 37°C

Advantages of colloid Fe(II)-Diethyldithiocarbamate [Fe(II)(DETC)₂]

• The only method for direct nitric oxide (NO) detection
• High efficiency of nitric oxide spin trapping
• Quantification of basal as well as stimulated NO production in aorta and vena cava
• Colloid [Fe(II)(DETC)₂] is "water-soluble" (easy to apply) and lipophilic (penetrates through membranes into cell layers)

EPR spectrometers suitable for this method

	MiniScope MS300, high sensitivity
	MiniScope MS100

Accessories required for this method

• Fixed temperature dewar for MiniScopes
• Holder which allows the adjustment of the altitude of the fixed temperature dewar in the cavity