localization - (Jun/23/2011 )
Hellow guys,
I am studying the localization of a protein. I transfected the Hela cells with a plasimd containing a RFP tag. The localization of my protein was showed in the attachement.
Do you guys have any ideas where my protein might locate? I have tried the marker for lysosome (Lamp-1), but it was suggested that my protein was not colocalized with Lamp-1. It would be nice that any of you can give me some suggestions about which marker I may use for the further detection. Thanks in advance.
Assuming that your cells are fibroblast looking, it looks like it's located rather exclusively in the cytosolic fraction. This is just a guess though.
lical208 on Fri Jun 24 02:53:11 2011 said:
Hellow guys,
I am studying the localization of a protein. I transfected the Hela cells with a plasimd containing a RFP tag. The localization of my protein was showed in tha attachement.
Do you guys have any ideas where my protein might locate? I have tried the marker for lysosome (Lamp-1), but it was suggested that my protein was not colocalized with Lamp-1. It would be nice that any of you can give me some suggestions that which marker I may use for the further detection. Thanks in advance.
I also would have thought lysosomes, but there seems to be too many spots inside the cells. Maybe mitochondria? Compare to click.
Looks to mitochondria to me...stressed mitochondria curl up: so how did you treat your cells? fixed, non fixed? You can use Mitotracker, it's a good dye.
Good luck
fysio lab on Fri Jun 24 08:38:16 2011 said:
Looks to mitochondria to me...stressed mitochondria curl up: so how did you treat your cells? fixed, non fixed? You can use Mitotracker, it's a good dye.
Good luck
Many thanks guys! I have fixed the cells. I also think about the mitochondria. But could early endosome be another option?
fysio lab on Fri Jun 24 08:38:16 2011 said:
Looks to mitochondria to me...stressed mitochondria curl up: so how did you treat your cells? fixed, non fixed? You can use Mitotracker, it's a good dye.
Good luck
Many thanks guys! I have fixed the cells. I also think about the mitochondria. But could early endosome be another option?
Yeah, mitochondria is a good option. You might want to try endoplasmic reticulum too.
lical208 on Fri Jun 24 02:53:11 2011 said:
Hellow guys,
I am studying the localization of a protein. I transfected the Hela cells with a plasimd containing a RFP tag. The localization of my protein was showed in the attachement.
Do you guys have any ideas where my protein might locate? I have tried the marker for lysosome (Lamp-1), but it was suggested that my protein was not colocalized with Lamp-1. It would be nice that any of you can give me some suggestions about which marker I may use for the further detection. Thanks in advance.
You could try using Mitotracker a marker specifically for mitochondria.Using sequential Confocal scanning look for co-localisationof the selected fluorophore.
Kindest regards.
Uncle Rhombus.