Application
IHC - Wholemount
Sample
Xenopus laevis Embryo (embryo stage 34)
Specification
embryo stage 34
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS 5% DMSO 20% BSA
Dilution
1/250
Secondary antibody
Name
Non-Abcam antibody was used: Monoclonal anti-rabbit IgG AP conjugate
Host species: Mouse
Clonality: Monoclonal
Conjugation: Alkaline Phosphatase
Host species: Mouse
Clonality: Monoclonal
Conjugation: Alkaline Phosphatase
Dilution
1/1000
Additional data
Additional Notes
1. Fix embryos in MEMFA for 20 min at RT
2. keep embryos in DENTS at -20 ¯C O/N (20% DMSO in methanol)
3. rehydrate embryos with stepwise 90%, 70%, 50%, 30% methanol, PBS (3 times)
4. blocking in 20% BSA in PBS (4-6h)
5. anti Sox2 in 20%BSA 5% DMSO PBS at 4¯C O/N
6. wash 2h PBS-TB (PBS, 0,05% Tween20, 0.2% BMB) (2 times) 2h PBS-TBN (PBS, 0,05% Tween20, 0.2% BMB, 0,3M NaCl) 5 min PBS-TB (3 times) PBS-TB O/N
7. 2nd antibody in 20%BSA 5% DMSO PBS (5h at RT)
8. wash 30 min PBS-TB (PBS, 0,05% Tween20, 0.2% BMB) (2 times) 30 min PBS-TBN (PBS, 0,05% Tween20, 0.2% BMB, 0,3M NaCl) 5 min PBS-TB (3 times) PBS-TB O/N
9. develop
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投稿 Oct 29 2014