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Author Topic: TH antibody  (Read 10990 times)

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Offline Annica Andersson

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TH antibody
« on: April 01, 2005, 07:35:38 AM »
Hi,
Does anybody know of a god tyrosine hydroxylase antibody (TH) for frozen rat tissue. I have tried TH from abcam with no satufactory result.
Annica

TH antibody
« on: April 01, 2005, 07:35:38 AM »

Offline richard03

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TH antibody
« Reply #1 on: April 01, 2005, 11:48:33 PM »
Annica,

Chemicon (Cat# AB152) TH works well on mouse and rat tissues.
Good luck.

Richard

Offline georgie

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help needed with chemicon TH
« Reply #2 on: April 05, 2005, 02:38:50 AM »
Hi

I've just started using the chemicon TH antibody on my frozen rat brain sections.  Although there is very clear specific staining, I'm having terrible problems with background.  If i run other antibodies in parallel, they have zero background.  So, my protocol and blocking steps do seem to be ok.

Anyone have any top tips?

Thank you
Georgie

Offline richard03

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TH antibody
« Reply #3 on: April 05, 2005, 09:21:50 AM »
TH antibody usually used very high dilution such as 1:2000~1:5000. What dilution you were using? You may need to adjust it.

Richard

Offline georgie

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TH antibody
« Reply #4 on: April 05, 2005, 09:40:25 AM »
Thanks for getting back to me, Richard.

I've tried the antibody at 1:2500, 1:5000 and 1:10000 and the staining was equally strong in each.  I'm now trying 1:20000, 1:40000 and 1:60000.  The crazy thing is that when i used to use the same antibody (i.e. same catalogue number but different batch), it was at 1:2000.  The recommended dilution in the data sheet is 1:2000-1:10000.

Tis a mystery to me....

Georgie :shock:

Offline richard03

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TH antibody
« Reply #5 on: April 05, 2005, 11:37:59 PM »
Sounds like the problem was not from primary Ab. You should check other factors such as 2nd Ab or blocking. Did you include a negative control?

Richard

Offline georgie

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TH antibody
« Reply #6 on: April 06, 2005, 02:59:04 AM »
I didn't use a negative control (not sure why - usually do).  However, I did use a positive control, in that I ran a different antibody that I've used successfully many times before.  The staining on that was beautiful.  Clear specific staining and zero background.

Thanks again for your reply  :)

Georgie

Offline richard03

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TH antibody
« Reply #7 on: April 06, 2005, 06:51:29 AM »
Is other antibody you used from same species as TH does? or did you use same 2nd antibody and detection?

Offline georgie

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All sorted
« Reply #8 on: April 20, 2005, 06:06:52 AM »
Well, its all working fine now.  Turns out it's a freakishly good antibody.  I'm now using it at around 1:40000 without nickel intensification.

Thanks for your help and suggestions
Georgie

Offline SGlerup

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Re: TH antibody
« Reply #9 on: December 06, 2012, 11:31:56 AM »
We use rabbit anti-TH from Pel-Freez (P40101) and from Millipore (AB152) in our lab.
Both antibodies are great for IHC on mouse and rat tissue.

We have reviewed them on www.pabmabs.com

Re: TH antibody
« Reply #9 on: December 06, 2012, 11:31:56 AM »