Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Embryoid bodies)
Specification
Embryoid bodies
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - triton-x100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Other product details
Dilution
1/80
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: 1% goat serum, 1mg/ml BSA, in PBS
Secondary antibody
Name
Non-Abcam antibody was used: anti-rabbit-488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
My cells are Bry-GFP ES cells and were at day 3.25 of EB differentiation, whereby ~90% cells were GFP (brachyury) positive by flow cytometry at staining. Therefore they should of stained positive but no staining was seen. NOTE: During the process of fixing and sectioning endogenous GFP signal marking Brachyury expression was lost as is common, allowing an anti-488 secondary to be used for staining.
Abcam response
This negative Abreview is currently being investigated by Abcam Technical Support. Please check here for an update response from Abcam.
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.
Abcam user community
Verified customer
投稿 May 28 2009