Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Pre-implantation mouse embryos (4 cell stage))
Specification
Pre-implantation mouse embryos (4 cell stage)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5 TritonX-100
Blocking step
BSA as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
Other product details
Dilution
1/250
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C
Secondary antibody
Name
Non-Abcam antibody was used: Alexa Fluor® 488 donkey anti-mouse IgG (H+L) *2 mg
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
4-cell stage mouse embryos were recovered and the zona pellucida removed. They were then fixed in 5% PFA for 20min at 37°C, permeabilized in 0.5% Triton X-100 (20min) and sequentially washed, three times, in PBS Tween-20 (0.1%) for 10min. Embryos were then blocked in 4% BSA for 4 hours at 4°C. They were incubated with ab67548 at 1:250 dilutions (in BSA) overnight. After three PBS Tween-20 washes and a further 4 hour BSA block, embryos were incubated in Alexa-fluor488 conjugated donkey anti-mouse secondary IgG for 1 hour at room temperature, washed a further three times in PBS Tween-20 and mounted onto coverslips in Vectashield + DAPI. Embryos were then imaged by fluorescence confocal microscopy and projected confocal stack images generated - as shown in two examples.
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. Alex Bruce
Verified customer
提交于 Apr 09 2009