Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (mouse brain (pons) (fixated, frozen and free-float)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Specification
mouse brain (pons) (fixated, frozen and free-float
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0,02% · Temperature: RT°C
Fixative
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0,02% milk in PBS-T
Dilution
1/100
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Invitrogen, A-11034
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
The image shows a mouse brain section, perfusion-fixated and only post-fixated 24 hours before cryoprotection in sucrose. Other material was included from brains that had been stored for +6 months in 1% PFA, giving no staining at all. It seems as if the epitope is sensitive towards over-fixation.
The samples were incubated with primary overnight at 4C, and on day two they were equilibrating for 1 hr at RT before washing and incubation with secondary.
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.
MRS. Trine Mikkelsen
Verified customer
投稿 Jan 11 2022