Hi!
I am during differentiation for SH-SY5Y cells. How can I prove that they really undergo differentiation? I can see some morphology changes and some neurites outgrowth, but I need some stronger proof. The best way would be WB or PCR for transcript changes. Have anyone tried something like this? Any suggestions, which proteins should I check?
Tau protein expression should increase with differentiation. You can also visually see the cells getting elongated with reduction in cytoplasmic volume.
Hi ,
I have worked on SK-N-SH cell lines.Once the cell line starts differentiating see if there is different in morphology. But,both matured and immatured cell lines usually look same.
If you want check if they have differentiated you can use WB or ELISA. Check for antibody which is expressed in matured neurons.
For SK-N-SH immatured cell lines, I have been using NCAM2 anti-rabbit antibody(cytoplasmic antibody) from sigma.Many companies provide both matured neuronal antibodies .Do ELISA or WB using matured and unmatured cells.Then you can see the difference.
You can also do PCR or immunofluroscence microscopy.
Good Luck!!!
Regards,
Pooja
Yeah I am going to use WB and check some neuronal markers, but I would like to know if anyone already tried it with some specific protein, marker and it worked? I mean you could see big difference between undifferentiated cells and differentiated?
Hi Anna, I am in the same boat as you.
Did you decide on a specific neuronal marker to check for differentiation? Did you find a significant difference between the undifferentiated and differentiated cells?
Can you advice?
I will try use anti-NSE ( neuron specific enolase) antibodies but don't know the results yet.
Hi Anna and Shruti, I am also doing the same thing. You can Neurofilament antibody. In my case it works very well..
Thanks Saranya and Anna...
I was wondering if we can also use MAP? Microtubule associated protein?
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