ICC: exotic problem with staining, vGlut1 ans synapsin-1
Posted 17 March 2011 - 04:52 AM
I have a problem with ICC on neurons, which I find rather exotic. I have tried to make a co-staining two synaptic markers, synapsin-1 (or synapsin-2 or synaptophysin) and vGlut1 (synaptic vesicle marker, receptor for glutamate), but I fail to see any staining. At the same time, when I stain with synapsin-1 only, I see specific bouton-like staining. I have not tried to stain with vGlut1 only, because I have just figured out that the problem comes from vGlut1 antibody. Could someone give me a clue why I can't stain two proteins at the same time or how to overcome this issue, sicne I want to see a colocalization! Thank you!
Posted 28 November 2011 - 07:32 PM
And do you use the same staining methods with these two antibody or two different ways, such as DAB and AP?
and then change the stain order. staining with vGlut 1 first.
Posted 02 December 2011 - 02:32 PM