Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Acetone
Permeabilization
Yes - 0.3% Triton X-100/PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Concentration
1 µg/ml
Incubation time
16 hour(s) and 0 minute(s) · Diluent: 0.5% BSA/PBS + 10 ug/ml goat IgG
Secondary antibody
Name
Non-Abcam antibody was used: Alexa Fluor 568 goat anti-chicken IgG
Host species: Goat
Clonality: Monoclonal
Conjugation: Alexa Fluor® 568
Host species: Goat
Clonality: Monoclonal
Conjugation: Alexa Fluor® 568
Concentration
2 µg/ml
Additional data
Additional Notes
This antibody binds quite nicely to glial cells not neuronal cells.
The images show coronal sections of adult mouse brain fixed with either acetone (top panel) or methanol (bottom panel). The field of view is the intersection between the cortex hemispheres, showing major blood vessels. Because this antibody is staining cells around blood vessels rather than neuronal cells within the cortex, it is likely that it is staining glial cells like astrocytes which are located around blood vessels and function to maintain the integrity of the blood-brain-barrier.
In serial sections were an isotype control (chicken IgY), was used instead of the ab75713 (images on the right), the staining was negative indicating that the pattern of staining with ab75713 is specific for astrocytes.
Compared with methanol fixation, the staining intensity for ab75713 is better after acetone fixation. When mouse brain sections were fixed in 2%PFA/0.05% gluteraldehyde, there was no positive staining observed with ab75713 antibody, even around major blood vessels.
The images show coronal sections of adult mouse brain fixed with either acetone (top panel) or methanol (bottom panel). The field of view is the intersection between the cortex hemispheres, showing major blood vessels. Because this antibody is staining cells around blood vessels rather than neuronal cells within the cortex, it is likely that it is staining glial cells like astrocytes which are located around blood vessels and function to maintain the integrity of the blood-brain-barrier.
In serial sections were an isotype control (chicken IgY), was used instead of the ab75713 (images on the right), the staining was negative indicating that the pattern of staining with ab75713 is specific for astrocytes.
Compared with methanol fixation, the staining intensity for ab75713 is better after acetone fixation. When mouse brain sections were fixed in 2%PFA/0.05% gluteraldehyde, there was no positive staining observed with ab75713 antibody, even around major blood vessels.
Abcam response
We would like to thank the reviewer for the time and effort they have taken to test this product and submit and Abreview. This product had not prevoiusly been tested in IHC-Fr on mouse.
For future experiments we would recommend trying a different fixative, such as paraformaldehyde and possibly an antigen retrieval step.
For future experiments we would recommend trying a different fixative, such as paraformaldehyde and possibly an antigen retrieval step.
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. Julie Nigro
Verified customer
提交于 Dec 28 2011