Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
No
Specification
Brain
Fixative
Paraformaldehyde
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS-Triton (0.3%) + azide (0.02%)
Dilution
1/100
Secondary antibody
Dilution
1/1000
Name
Non-Abcam antibody was used: Alexa Fluor® donkey anti-mouse
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 568
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 568
Additional data
Additional Notes
The tissues were fixed (animals perfused fixed) with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.
Mouse was injected with a virus expressed specificaly in astrocytes. The range of dilution used was 1:100 to 1:3000 (1:100, 1:300, 1:1000 and 1:3000). The picture was taken in the site of injection. The staining obtained with or without primary antibody are similar. The observation is only the fluorescence of the mCherry protein. Images were taken with the same settings on the microscope.
Didn't work on mouse brain sections.
Abcam response
Many thanks for your valuable feedback. ab125096 has not been validated for use in this application. It appears that under these experimental conditions, the antibody does not enhance the levels of fluorescence observed with endogenous mCherry. We welcome further customer feedback. For more information, please contact scientific support.
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. Karine Thibault
Verified customer
提交于 Nov 04 2014