User:Mar/Notebook/2007-8-15

= Mouse embryos E16 injection with BrdU =

Goal

 * 1) Assesment of BrdU-labeled cells' migration during undisturbed ontogenesis
 * 2) To be co-immunostained for BrdU and nestin

Animals
Mothers: rl104b - 34g rl109e - 28g

Embryos:

Materials

 * BrdU solution: 5mg/ml in saline with 0.007 N NaOH (i.e. dilute 2N NaOH 286x). NaOH usually unnecessary.

Procedure
Protocol follows the original experiment. BrdU injected to a pregnant mother:
 * dose 50µg/g
 * single injection labels during period extending from 15min to at least 2hr

Injection regime: 34g and -> inj'd 0.34mL-45'-0.34mL 28g and -> inj'd 0.28mL-45'-0.28mL

i.e. total volume 2x.34+2x.28=1.24mL Make 1400µL from 1.4*5mg = 7mg

= Western with John T (1) = PVDF casein blocking