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FasL? - I need to activate Fas apoptotic pathway... (Jul/27/2005 )

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Hi,
I've got 2 drugs we're certain are acting by interferring with the Fas apoptotic pathway ONLY.

We need to check this by replacing one of these drugs with another Fas apoptotic pathway specific agent.

I thought I could just "buy" some fas ligand but I can only find antibodies for it, not the ligand itself.

Does anyone know if one can buy Fas Ligand or know of a drug that is cytotoxic specifically (or moderately specifically smile.gif ) through the Fas/FADD apoptotic pathway?

Thanks

-Mummytomax-

sigma-aldrich have FasL from humans and mouse:

http://www.sigmaaldrich.com/

Fas Ligand from Mouse (1)
F0552 >95% (SDS-PAGE), recombinant, expressed in mouse NSO cells, lyophilized powder (Sigma)

Fas Ligand human (1)
F0427 >95% (SDS-PAGE), recombinant, expressed in CHO cells, lyophilized powder (Sigma)

Fas Ligand human (1)
F4428 >95%, recombinant, expressed in HEK-293 cells (amino acid residues 133-281, N-terminal FLAG tagged) (Sigma)

Fas Ligand, soluble human (1)
S8689 recombinant, expressed in human embryo kidney cells, lyophilized powder (Sigma),

for example.

mike

-jadefalcon-

Right - the obvious sorce is NEVER the one you check LOL!!

Had managed to find one other company but it was in 'reserach production stage only"

Thanks

-Mummytomax-

try with
SuperFasLigand™, Soluble (human) (recombinant)
http://www.axxora.com/APO (searching for FasL)

Seb_

-tryptofan-

Yeah I agree

SuperFasL works the best in our hands. Some of the commercial FLAGFasL constructs require cross linking with anti-FLAG antibodies to achieve the level of activity that SuperFasL has.


Good Luck

-Mountainman-

QUOTE (Mountainman @ Sep 20 2005, 05:28 PM)
Yeah I agree

SuperFasL works the best in our hands. Some of the commercial FLAGFasL constructs require cross linking with anti-FLAG antibodies to achieve the level of activity that SuperFasL has.


Good Luck


Hi guys, I have placed an order for SuperFasL as a positive control. I am trying to find out if the FASL is present in my CHO cell system.
Does anybody know if the FASL is phosphoprotein?
I also have a doubt if the presence of FAS in cells always confirms that FASL also must be present.
Please have your views, to clear my doubt.
Thanks

-indianajones-

QUOTE (Mummytomax @ Jul 27 2005, 11:45 PM)
Hi,
I've got 2 drugs we're certain are acting by interferring with the Fas apoptotic pathway ONLY.

We need to check this by replacing one of these drugs with another Fas apoptotic pathway specific agent.

I thought I could just "buy" some fas ligand but I can only find antibodies for it, not the ligand itself.

Does anyone know if one can buy Fas Ligand or know of a drug that is cytotoxic specifically (or moderately specifically smile.gif ) through the Fas/FADD apoptotic pathway?

Thanks


no no, you can simply buy anti-fas L antibody and trigger apoptosis. the antibody works as a fas ligand, no worries. there are many papers about it. This is a famous method for triggering Fas pathway, called Fas-induced apoptosis

read the attached file.

-Curtis-

Hi Guys
Does anybody work on FASL from CHO cells??? I wanted to know if the FASL of CHO origin is phosphoprotein.
Thanks!!!

-indianajones-

QUOTE (indianajones @ Sep 18 2008, 12:42 AM)
Hi Guys
Does anybody work on FASL from CHO cells??? I wanted to know if the FASL of CHO origin is phosphoprotein.
Thanks!!!


I have published a couple papers on FasL during my masters. I would say it is possible that FasL intracellular domain is phosphorylated (I always suspected it...), but there is to my knowledge no documented evidence that FasL is a phosphoprotein. You can PM me if you want to discuss it further. I have no conflict of interest since I no longer study FasL and my old lab shut down anyways.....


-Mountainman-

QUOTE (Mountainman @ Oct 2 2008, 07:16 AM)
QUOTE (indianajones @ Sep 18 2008, 12:42 AM)
Hi Guys
Does anybody work on FASL from CHO cells??? I wanted to know if the FASL of CHO origin is phosphoprotein.
Thanks!!!


I have published a couple papers on FasL during my masters. I would say it is possible that FasL intracellular domain is phosphorylated (I always suspected it...), but there is to my knowledge no documented evidence that FasL is a phosphoprotein. You can PM me if you want to discuss it further. I have no conflict of interest since I no longer study FasL and my old lab shut down anyways.....


Hi Dear
Thanks for your post against my querry.
Actually I am trying to explore if FASL is present into my CHO cell system. I ran western blot sufficient number of times, but unsucessful. I know its a transmembrane protein. My impression is, the lysis buffer which I am using is sufficient to get whole cell extract. I am not sure what is the problem if its lysis buffer problem or SDS-PAGE.
Could I be helped in this case??
Thanks!!!!

-indianajones-

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