User:Mar/Notebook/2007-8-15
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Mouse embryos E16 injection with BrdU
Goal
- Assesment of BrdU-labeled cells' migration during undisturbed ontogenesis
- 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 [1]. 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