Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Trigeminal ganglion)
Permeabilization
Yes - 1% triton, 30 minutes
Specification
Trigeminal ganglion
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: Normal Donkey Serum
Dilution
1/5000
Secondary antibody
Name
Non-Abcam antibody was used: Donkey anti-Chicken Alexa 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
We performed immunohistochemistry on the trigeminal ganglion to test expression of mCherry. We used a AAV serotype 8 virus expressing mCherry as a reporter (AAV-CMV-mCherry). The expression of mCherry was detected in some sensory neurons (trigeminal ganglion). We used anti-mCherry antibody and secondary with a different channel (Alexa-488) and observed complete overlap of anti-mCherry immunoreactivity with natural fluorescence of mCherry reporter. Strong mCherry signal was detectable with 1:5000 dilution and higher concentrations resulted in increased background levels.
In addition we verified this antibody using a transgenic mouse expressing mCherry in a subset of neurons (unpublished data, picture not included), and observed complete overlap of anti-mCherry immunoreactivity with the reporter mCherry.
We conclude that this antibody is useful for the detection of mCherry even at low levels of expression and show very specific staining. We suggest to use 1:5000 or even higher dilution for immunohistochemistry experiments.
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. Guennadi Dvoriantchikov
Verified customer
提交于 Feb 08 2018