Kinase screening

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Nicole
Nicole's picture
Kinase screening

Compounds that control kinases activity have a great interest in drug development. Kinases are enzymes that catalyze phosphorylation and therefore are involved in different diseases such as cancer. Scientists have used different tools to screen kinases such as kinase profiling by radioactive assay or fluorescence-polarization method. I have heard that kinase inhibitors fail most of the time to deliver what they claim. Does anyone know names of companies and test kits used for kinase screening? also kits that track signal-transduction pathways, I need to set up an experiment to observe the effect of an antibody in signal-transduction within the cell. Are any products available to observe these activities inside the cell?

Larkia
Larkia's picture
Well, I work at the in-vitro

Well, I work at the in-vitro kinase level, not the in-cell level, so I can't help much with the in-cell products, but I know of a number of test kits for kinase screening:

Upstate offers a number of KinEASE (FP) kits for various kinases. Invitrogen offers both Z-Lyte (FRET) kits and LanthaScreen (TR-FRET) technology. Molecular Devices has IMAP (FP) technology. BioSource offers various phosphoELISA kits for a number of kinases.

For tracking signal-transduction pathways, you may want to check out Kinexus. They have various pathway screens, though I believe you have to send them your sample(s).

king of WB
king of WB's picture
So you may help me with this!

So you may help me with this! I am using an in vitro system too. I am studiing an antiinflammatory drug and I found that this drug inhibit nfkb activation and prevent Ikba phosphorylation and degradation. Now I am about to see what goping on on upstream signal. So doing IKK assay. I was reading all I can find articles/protocols...and I am confused because some people used IKKa or IKKb or IKKg to immunoprecipitate the protein from the lysate. Could any one have ptotocol for IKK assay and which antibody should I used

samm
samm's picture
IkBa is most commonly used -

IkBa is most commonly used - its most imp as its perinatal lethal in KO mice. IKK is however more variable - you may want to check for your ap.
You might want to check out this reference for a coordinated model for all the isoforms - its somewhat old, but well written.
Science, Vol 298, Issue 5596, 1241-1245 , 8 November 2002
[DOI: 10.1126/science.1071914]

Elisa
Elisa's picture
Aside from using NFkB siRNA

Aside from using NFkB siRNA or NFkB-/- cells/mice to show downstream signaling to IFkb, there are commercial assays available at

http://www.cellsignal.com/index.asp?cookie%5Ftest=1

or

http://www.upstate.com/home.q./Upstate+Cell+Signaling+Solutions

Signma Aldrich also has an extensive line of products for kinase signal transduction activity assays.

Hope this helps.