Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Other product details
Dilution
1/100
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS + 0.3 % Triton X100
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit Alexa Fluor 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
I used this antibody on sections of rat brain (30 um) in free floating.
I did not observe any nuclear staining as it was expected. Instead, I observed some staining in glial cells that looks like oligodendrocytes. There is a punctuated and diffuse staining too.
The pictures loaded show the staining I observed in the cerebral cortex (A) using the antibody diluted 1:100, and show the negative control (B) (i.e. sister incubated with the diluent, but without any primary antibody). Note that the images in A and B were taken using the same set-up of acquisition.
The tissues are from perfused fixed animals perfused with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.
I did not observe any nuclear staining as it was expected. Instead, I observed some staining in glial cells that looks like oligodendrocytes. There is a punctuated and diffuse staining too.
The pictures loaded show the staining I observed in the cerebral cortex (A) using the antibody diluted 1:100, and show the negative control (B) (i.e. sister incubated with the diluent, but without any primary antibody). Note that the images in A and B were taken using the same set-up of acquisition.
The tissues are from perfused fixed animals perfused with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.
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.
Dr. Sophie Pezet
Verified customer
投稿 Jun 17 2010