What assays to use for the following... - Angiogenesis, proliferation, apoptosis...... (Sep/19/2008 )
Hi all,
I am currently working on a therapy for cancer. We are targeting a protein that is involved in 80% of cancers. It is supposed to contribute in angiogenesis, evading apoptosis, proliferation and suppress the immune system. I would like to perfom a series of assays for this target molecule. Can anyone please suggest me
1. A cost-effective and the best in vitro and in vivo angiogenesis assay kits available.
2. How to go about testing if this molecule is an indirect suppressor of immune system
3. A good proliferation assay (currenlt i am using MTT proliferation assay, but would liek to test it using other methods as well)
4. A good apoptosis assay kit. - we did try few apoptosis methods but somehow we r not able to get it right. So if anybody can suggest an apoptosis assay with their protocol would be fantastic.
Thank you all very much.
I am currently working on a therapy for cancer. We are targeting a protein that is involved in 80% of cancers. It is supposed to contribute in angiogenesis, evading apoptosis, proliferation and suppress the immune system. I would like to perfom a series of assays for this target molecule. Can anyone please suggest me
1. A cost-effective and the best in vitro and in vivo angiogenesis assay kits available.
2. How to go about testing if this molecule is an indirect suppressor of immune system
3. A good proliferation assay (currenlt i am using MTT proliferation assay, but would liek to test it using other methods as well)
4. A good apoptosis assay kit. - we did try few apoptosis methods but somehow we r not able to get it right. So if anybody can suggest an apoptosis assay with their protocol would be fantastic.
Thank you all very much.
MTT is one of the best proliferation assays about, just not very sensitive at the low range. I guess you could do a DNA content assay of some sort to give growth curves.
Apoptosis: Caspase 3 is the gold standard assay, but you could western or ELISA for cleaved PARP.