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"A number of imaging markers which target hypoxia exist and are widely used in research. EF5 is a compound developed at the University of Pennsylvania by Dr. Cameron Koch and Dr. Sydney Evans. Upon injection into animal tissues, EF5 selectively binds to hypoxic cells and forms adducts. A mouse monoclonal antibody, clone ELK3-51 which is directly conjugated to Cyanine 3 is then used to selectively bind the EF5 adducts. Advantages of the EF5 Hypoxia Detection Kit:. EF5 exists in only one form:Pimidozole, an alternative hypoxia marker, exists in two forms, one of which is charged and very hydrophilic, the other lipophilic. Pimidozole thus has a very complex biodistribution. In contrast, EF5 is lipophilic and uncharged and this allows very rapid and even tissue distribution. . EF5 binding images can be calibrated to provide quantitative data on the pO2 values of each cell (Koch CJ, 2002):The fluorescent images obtained from EF5 binding can be calibrated according to camera settings and a ""cube-binding"" value which is obtained through a separate procedure. The intensity values of calibrated images are directly related to actual tissue pO2 values. As a result, these images provide information regarding not only where hypoxic areas may or may not be, but also data regarding the distribution and levels of hypoxia. Reference:Koch CJ (2002) Measurement of absolute oxygen levels in cells and tissues using oxygen sensors and 2-nitroimidazole EF5. Methods in Enzymology 352: 3-31."