Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (E13.5 placenta)
Permeabilization
Yes - 5% goat serum and 0.05% Triton X-100 in PBS
Specification
E13.5 placenta
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde
Other product details
Incubation time
8 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% goat serum and 0.05% Triton X-100 in
Dilution
1/1000
Secondary antibody
Dilution
1/700
Name
Non-Abcam antibody was used: Goat anti Rabbit Alexa Fluor 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Additional data
Additional Notes
Placentas at E13.5 were fixed in 4% PFA overnight, washed in PBS and embedded in OCT for cryosectioning.
Sections were blocked and permeabilized in blocking solution: 5% goat serum and 0.05% Triton X-100 in PBS for 20 min.
Sections were incubated with the primary antibodies over night at 4 degrees in blocking solution. Primary antibodies were ab104401 (1/1000 dilution) and anti GFP (Developmental Studies Hybridoma Bank). GFP labels the embryonic placental vasculature in our mouse model.
Sections were washed with PBS and incubated with the secondary antibodies goat anti rabbit Alexa Fluor 594 (red), and anti mouse Alexa Fluor 488 (green) in a 1/700 dilution in blocking solution.
Sections were washed with PBS and mounted in Vectashield mounting medium with DAPI.
This staining revealed a specific signal for ab104401 in spongiotrophoblast (red).
The image represents the central portion of a cross section of a placenta at embryonic day 13.5. The decidua, spongiotrophoblast and labyrinth layers are indicated.
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. Paul Delgado Olguin
Verified customer
投稿 May 13 2015