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Hi,
I plan to do a colony foration assay (or foci formation assay) to observe the effect of my plant extract, of certain concentration, on cancer cell lines. I plan to seed about 300-500 cells per 60mm dish and observe the no. of colonies that form after about 2 weeks. In these two weeks, can I change medium (containing the right dosage of the extract)?
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colony formation assay
19 May 2013 - 11:29 PM
5'-azacytidine treatment
24 April 2013 - 11:57 PM
I'm treating my cell lines with 5ázacytidine to check for methylation. I'm planning to treat my cell lines with 5ázacytidine for 5 days. Do I have to replace the medium and drug every day or can I do it alternatively?
Álso, Ive made my stock solution of 5ázacytidine (Sigma, A2385) in DMSO and frozen the aliquots in
-80 degrees.
Thank you
Álso, Ive made my stock solution of 5ázacytidine (Sigma, A2385) in DMSO and frozen the aliquots in
-80 degrees.
Thank you
random primers or oligodT
20 March 2013 - 01:05 AM
I'm confused as to use random or oligodT primers. I have to check expression of various egenes in around 130 tissues. Most reasonable kits (with 100+ reactions), use randome hexamer primers.
I checked my 12 genes of interest on cancer cell lines whose RNA was extracted using olidodT primers. I would ideally like to use oligodT primers for my RT-PCR in the clinical tissues, but the kits are very expensive. The cheaper kits use random primers.
Any advice?
I checked my 12 genes of interest on cancer cell lines whose RNA was extracted using olidodT primers. I would ideally like to use oligodT primers for my RT-PCR in the clinical tissues, but the kits are very expensive. The cheaper kits use random primers.
Any advice?
study of receptors in cell lines
15 March 2013 - 01:29 AM
Hi,
I'm styudying receptor in cancer cell lines. Since it is overexpressed I plan to observe the effect of a receptor antagonist in the cancer cell lines (e.g. MTT by proliferation). My problem is that the antagonist that's available may not be 100% selective to the recetor I'm studying (e.g receptor A) and that could also affect receptor B as well.
How do I ensure that the B receptor -selective antagonists are not active in my assay to have more compelling results.
I was thinking I treat the cell lines with antagonist to receptor A alone, antagonist to receptor B alone and combination of both antagonists of A and B?
I'm styudying receptor in cancer cell lines. Since it is overexpressed I plan to observe the effect of a receptor antagonist in the cancer cell lines (e.g. MTT by proliferation). My problem is that the antagonist that's available may not be 100% selective to the recetor I'm studying (e.g receptor A) and that could also affect receptor B as well.
How do I ensure that the B receptor -selective antagonists are not active in my assay to have more compelling results.
I was thinking I treat the cell lines with antagonist to receptor A alone, antagonist to receptor B alone and combination of both antagonists of A and B?
chemical-induced HCC mouse model
28 January 2013 - 11:09 PM
Hi,
I've never worked with mouse models but have to plan for one and would appreciate any advice.
I looking to develop a mouse model that forms HCC after cirrhosis. I've read a few papers and most use CCL4. Has anyone used this treatment and if so:
1. howl ong doe sit to take to develop cirrhosis
2. how long doe sit take to develop HCC following cirrhosis
3. the % of survical of mice. Can teh mice survive til the development of HCC
4. Is this a harsh treatment?
Many thanks
I've never worked with mouse models but have to plan for one and would appreciate any advice.
I looking to develop a mouse model that forms HCC after cirrhosis. I've read a few papers and most use CCL4. Has anyone used this treatment and if so:
1. howl ong doe sit to take to develop cirrhosis
2. how long doe sit take to develop HCC following cirrhosis
3. the % of survical of mice. Can teh mice survive til the development of HCC
4. Is this a harsh treatment?
Many thanks
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