Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Antigen retrieval step
None
Permeabilization
Yes - 0.3% Triton-X 100
Specification
Brain sections
Blocking step
(agent) for 45 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Other product details
Incubation time
0 minute(s)
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: anti-Rabbit antibody
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Additional data
Additional Notes
The abreview is for IHC-FoFr (free floating sections)
The sample was sectioned after being fixed with 4% PFA and then frozen to be sectioned on a microtome (30um sections). We tried many concentrations of the primary and tried to permeabilize using 0.3% Triton-X 100. The signal is not high enough above background to distinguish cells containing SIRT1.
The following describes the image attached:
(A): Darkfield illumination of the tissue to show tissue architecture. The visualized region is the pyramidal cellular region adjacent to the alveus.
(B): Same region illustrated by (A) visualized by a red filter to illuminate the AlexaFluor 594 secondary antibody.
(C): Same region illustrated by (A) and (B) visualized by a green filter to visualize any nonspecific fluorophore fluorescence.
(D): Pyramidal cells published by Zachary S. et. al. (PMCID: PMC3071026) to show positive SIRT1 immunoreactivity.
This SIRT1 antibody (ab32441) is not labeling rat SIRT1.
Abcam response
Thank you for submitting this review as part of our AbTrial Testing Program. Your results, in additional to our preliminary results, suggest that ab32441 is not reactive with rat species. We welcome additional Abreview feedback.
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.
Vaibhav Konanur
Verified customer
提交于 Mar 09 2016