User:Mar/Notebook/2007-8-15

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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 [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

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