Re: Querry regarding BrdU

From: vivek tanavde (vtanavde@hotmail.com)
Date: Fri Nov 02 2001 - 22:44:54 EST



Hi Preeti,

The APO-BRDU kit available from Pharmingen should work for you. You should be able to get a horse shoe shaped profile on a dot plot of BrDU on the y-axis v/s PI on the x-axis. The apoptotic cells will be sub G0/G1 and negative for BrDU. We have looked at the effect of tyrosine kinase inhibitors on human myeloid cell lines and we were able to see a G0/G1 arrest, followed by most of the cells undergoing apoptosis upon longer incubation with the drug using this assay.


Vivek Tanavde

Johns Hopkins Comprehensive Cancer Center
Bunting-Blaustein Cancer Research Bldg, Room 268
1650 Orleans Street
Baltimore, MD 21231


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attached mail follows:


I want to place one querry in front of the forum.I hope that the forum will help me out.
I want to know that if I'm getting cell cycle arrest at one dose of any chemotoxic agent & I want to see that at any other dose I'm getting increased apoptosis,Can I couple the two techniques of BrdU labelling & Propidium iodide staining to see this effect.Or more intricately,If I want to demarcate the apoptotic fraction  precursor population(as being S phase arrested or G2-M phase arrested) ,how can I do this.
Or how can I know that which of population is going towards apoptosis(G0-G1 arrested or G2-M arrested population).
If somebody can help telling this,please reply me as soon as possible.
Thanx in advance.
Preeti
PhD student
India 


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