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Post subject: Triple Staining with TUNEL+PI+Cardiomyocytes
Hi Everyone,
Thanks for the earlier reply.
I am back again to Immunohistochemistry. I know Carl doesn't like TUNEL.
Recently I did TUNEL staining in cardiac section and now i want to prove that origin of apoptotic cells are from heart section.
I am planning to use TUNEL+PI+anti smooth muscle antibody - Fluorescent staining. I found this technique in some journal.
Is there better recommendation to this.
Thanking you all in advance.
Milan
Fri Feb 04, 2011 8:46 am
Carl
Rank: Member
Joined: Oct 11, 2004
Posts: 1871
Post subject: welcome back, milan
lol....sure, TUNEL is a good tool in Good Hands
So, you want to prove that a TUNEL-positive cell is an apoptotic myocyte in your cardiac muscle section?
I do not know what "PI" is.
Also, why would you use anti SMA?
I may be wrong again but, an "anti smooth muscle antibody " will not demonstrate cardiac myocytes?
Sure, the blood vessel smooth muscles will be positive.
Curiously,
Carl
Sat Feb 05, 2011 5:14 pm
milan
Rank: Member
Joined: Nov 04, 2010
Posts: 16
Post subject:
Dear Carl,
Thank you for your reply as always.
I am sorry that i mentioned Anti smooth muscle antibody, I must be thinking something.... my bad.
Well, I planned to use anti Desmin or anti alpha sarcomeric actin or any other to show the presence of cardiomyocytes.
PI = Propidium Iodide staining.
My plan is
1. Stain the section by TUNEL = green fluorescence.
2. Stain by PI ( which will stain all nucleus red ) and apoptotic cells will be stained yellow.
3. Then finally stain with fluorescent labelled above antibodies to determine the location.
I found some journals explaining this procedure.
My aim is just to detect the origin of apoptotic cells, later on i will be using qPCR or gene array to determine more. I will be looking at some apoptotic signals too.
Please advise me better options.
Thanks
Milan
Wed Feb 09, 2011 1:14 am
milan
Rank: Member
Joined: Nov 04, 2010
Posts: 16
Post subject:
Hi again,
I want share some more information which i learnt.
The link below explains how triple staining was done in cardiac section.
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