Evan's Blue 0.5% in PBS

Inject 200 µl 0.5% Evans Blue/PBS (1 mg) iv

Observe mouse for 30 minutes

Euthanize, collect & weigh tissues. Cervical dislocation alone preferred since this causes less  change in EC premeability than other methods such as CO2 narcosis or drug overdose.

Add 500 µl formamide

55˚C 24-48 hr.

Assay A600 (blank is 500 µl formamide, EB/formamide as standard curve)

Present data as extracted ng Evans Blue/mg tissue

Ref: Ref: J Vis Exp. 2013 Mar 16;(73):e50062. doi: 10.3791/50062.