ICC: exotic problem with staining, vGlut1 ans synapsin-1 - (Mar/17/2011 )
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!
first you said there is a problem about vGlut1 antibody, maybe the antibody affect the result. first use the correct antibody.
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.
If you could post details of your protocol, we may be able to help further.