Application
IHC - Wholemount
Sample
Mouse Embryo (Wholemount immunostaining of E6.5 mouse embryo)
Specification
Wholemount immunostaining of E6.5 mouse embryo
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS/0.1% Triton-X/5% donkey serum/1% BSA
Dilution
1/100
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Donkey anti rabbit Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
Good specific staining and low background. Embryonic day E6.5 and E7.5 embryos were tested and staining looked good on both.
Immunostaining protocol:
1. Embryos were fixed for 15 mins at room temperature in 4% PFA.
2. Embryos were permeabilized for 30 mins at room temperature in PBS + 0.5% Triton-X.
3. Embryos were blocked overnight at 4 degrees C in PBST (PBS + 0.1% Triton-X) with 5% donkey serum and 1% BSA.
4. Embryos were incubated overnight at 4 degrees C with primary antibodies diluted in blocking buffer. All Abcam antibodies were tested at 1:100 dilution.
5. Embryos were washed 3x for 15 mins in PBST and blocked for 3 hours in blocking buffer.
6. Embryos were incubated overnight at 4 degrees C with secondary antibodies (Alexa Fluors) diluted 1:500 in blocking buffer.
7. Embryos were washed 3x 15 mins in PBST and stained with a nuclear dye (Hoechst).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Sophie Morgani
Verified customer
提交于 Feb 09 2018