I'm looking at some protein tyrosine phosphatases that I express transiently in HEK293/293T cells. The extracellular domains of these proteins are thought to be highly n-glycosylated. Is there a way to visualise this glycosylation using confocal microscopy?
I agree with Diluka, but if your cells are formalin fixed and paraffin embedded you can use the lectin histochemistry tecnique. There are many pulications about this technique
Good luck
But that would be non-specific detection? it will show me all the proteins that are n-glycosylated not just the proteins of interest? and will it stain the membrane too?
Thanks
I understand, if you want to be more specific in the detection you could try to work at protein level. May be you could obtain a membrane fraction of your cells in order to achieve the electrophoretic separation of the membrane proteins, transfer these proteins to nitrocellulose or PVDF membranes and detect them using lectins-HRPO conjugates (direct method) or lectins and secondary antibodies-HRPO against the lectins (indirect method). As I am sure that you know the MW of your protein and probably you have antibodies against the protein, you could easily visualise
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