Topics » Signaling Pathways

PI3K activity

We would like to determine PI3K activity in monocytes. Could anybody help us with the methodology? Or, alternatively - is there any available kit that works fine?
Thank you

Maria-Rosa Sarrias ⋅ Feb 23, 2012
10 Replies
  • Carlos Guillen

    Hi. I determined the PI3 kinase activity by a radioactive method. It was published in Endocrinology (Guillen C et al. Endocrinology 2006). I hope that i can contribute to your work. However, there are some methods not using radioactivity. So, as it has been suggested by other people you could use it. In contrast, the sensitivity could be lower that that with radioactivity. Good luck!!. Carlos

    Mar 7, 2012
  • Marinaldo Neto

    Hello Maria Rosa,

    Not yet tested in my experiments because the orders have not arrived, but before ordering I looked and saw the kit being used in the methodology of some papers and I saw some comments about the kit and I seemed to be very interesting to test, mainly because of using an ELISA technique as accessible and do not use radioactive labeling. I asked the technical manual to the company, if you want I can send it via email and you analyze and see if it's interesting for their experiments.

    I hope I have helped

    Mar 5, 2012
  • Maria-Rosa Sarrias

    Hello,
    thank you very much to all of you. Marinaldo, did you try the kits you recommend?
    Thanks again!

    Mar 5, 2012
  • Emma Amir-Ebrahimi

    Thanks Marinaldo. I'm focusing on Class II and am struggling to understand all the signalling pathways!

    Mar 3, 2012
  • Marinaldo Neto

    Hello Emma

    The kit that I mentioned above, using ELISA, it measures the function of Class I PI3K

    Mar 3, 2012
  • Wenjun Zhou

    WB is a regular method for investigating the PI3K pathway. Phosphorylation or dephosphorylation of their downstream proteins should be focused.

    Mar 3, 2012
  • Emma Amir-Ebrahimi

    Hi Guys

    Hope this isn't a silly question, but are you all looking at Class I PI3K?

    Mar 3, 2012
  • Yonara Marques

    Dear Maria Rosa,

    I also study this pathway and I just want to suggest you to include PTEN protein in you study, because this one is really important in the PI3K pathway. You can study by WB or Elisa assay.
    Good luck

    Mar 2, 2012
  • Marinaldo Neto

    Hello Maria Rosa! Recently I also came researching possible tests to evaluate the function of PI3K. As Laura said, there are some in vitro assays using antibodies labeled with radioactive substrates, if I remember correctly, labeled with europium, which prevented me from buying it. Then I found an assay that measures the function of PI3K by an ELISA assay, called "PI3-Kinase Activity ELISA: Peak", as well as a kit for quantification of PIP3 called "PIP3 Mass ELISA kit," both the company Echelon.

    Thanks,
    Good luck and good results

    Mar 2, 2012
  • Laura Braccini

    To determine the Pi3k activity you should use an in vitro assay termed lipid kinase assay. However this assay use radioactive substrate and it is not simple to do, thus I believe that you could perform a simple Wb assay against the phosphorylated form of Akt. It is a well known in vivo downstream effector molecule of PI3K.

    I hope that my mail is usefull for you

    Good luck

    Feb 28, 2012
Follow this Post

Contributors

Share