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Latrunculin A and cytokine treatment in the same time

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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Latrunculin A and cytokine treatment in the same time

Postby roxanalibe » Tue Jul 13, 2010 9:16 am

Hallo to all of you,

I am the first time writing to this group and I am happy to do that, as I would like to network with more people dealing with my biology/biochemistry topics.

Context: So, shortly, I need to treat transiently transfected cells (in a dual luciferase assay) with cytokine for a certain receptor and asses the signaling level in time, +/- Latrunculin A (a cell permeable marine toxin that disrupts microfilament organization by formation of a 1:1 complex with monomeric G-actin). As I believe there is an involvement of the actin in the signaling via the receptor I study, I am trying to see that using this approach.

Question: could entry of the Lat A interfere with the binding of the cytokine to the receptor? Is it possible to treat the cells in the same time with both compounds and not affect the effect of each of them?

Thank you in advance,
Roxana
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Postby kolean » Wed Jul 14, 2010 3:13 am

Would you treat the cells with the Latrunculin A first, to bind up all the monomeric G-actin first, and then treat the cells with the cytokine? That way the momomeric G-actin is all bound up and can not participate in the signaling of the cytokine, and you can asses the signaling with the dual luciferase assay.
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Postby roxanalibe » Wed Jul 14, 2010 7:53 am

thanks a lot, this is useful and fast, as i will begin the treatment in one hour
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