I would like to detect motor neurons in mouse spinal cord as an alternative to violet crystal.
Hello,
we use an antibody against Choline-Acetyltransferase from Millipore developed in goat (A-ChAT, gt, Millipore) and for immunofluorescence we dilute it 1:400 when afterwards detected with secondary antibodies conjugated with red fluorescent CY3 (Anti-goat FaB-Fragments developed in mouse from Jackson).
In DAB stainings we use a dilution from 1:1000, followed by detection with a Vector-Kit.
However, if Dr. Schmidt provides you a free test sample, it is worth to test it!!
Sincerely Yours
Stefan
Hi Christina.
I'm working for the company Synaptic Systems. We have just recently developed a new antibody against ChAt (Choline-Acetyltransferase), here is the link;
https://www.sysy.com/products/chat/facts-297013.php
It works well in IHC and IHC-P in mouse tissue. Please contact me at [email protected] if you want to test a free sample.
Best regards,
Carsten
Hello,
we use an antibody against Choline-Acetyltransferase from Millipore developed in goat (A-ChAT, gt, Millipore) and for immunofluorescence we dilute it 1:400 when afterwards detected with secondary antibodies conjugated with red fluorescent CY3 (Anti-goat FaB-Fragments developed in mouse from Jackson).
In DAB stainings we use a dilution from 1:1000, followed by detection with a Vector-Kit.
However, if Dr. Schmidt provides you a free test sample, it is worth to test it!!
Sincerely Yours
Stefan
This has been asked several times on RG. Most attest to the goat polyclonal antibody from Milipore (AB144P) that Stefan mentions.
We use it in mice, rats, and turtles.
- Incubate O/N in the fridge with primary ChAT antibodies (goat anti-ChAT antibodies (1:250, Milipore, USA) in blocking buffer.
Incubate O/N in fridge with secondary Alexa488 conjugated to donkey anti-goat antibody (1:333, Jackson)
Works every time.
In mouse: http://www.jneurosci.org/content/35/8.cover-expansion
In turtle: http://www.jneurosci.org/content/34/8.cover-expansion
The above images are from 100um slices imaged using a confocal.
We have also used it to stain turtle spinal cord segments treated with CLARITY.
http://www.emdmillipore.com/US/en/product/Anti-Choline-Acetyltransferase-Antibody,MM_NF-AB144P
Hello Christina,
upvoted?
Therefore, I think the Millipore-AB is working?
Am I right?
Sincerely Yours
Hello Stefan,
The first I tested was AB143 from Millipore and I have found difficulties to detect ChAT because it appears to much DAB unspecific signal. I must test other options such cryosection instead paraffin included tissue and the antibody free sample provided by Carsten. Than you for your interest and I would appreciate so much to have some technical help.
It seems this thread is quite old now, but could anyone comment on whether it is necessary to first perfuse the mice before performing a staining using the Millipore ChAT antibody? Thanks!
- Badges
- Science method