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#1 lical208

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Posted 23 June 2011 - 06:53 PM

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.

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Edited by lical208, 24 June 2011 - 03:16 AM.


#2 zienpiggie

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Posted 23 June 2011 - 10:18 PM

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.

#3 Rsm

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Posted 24 June 2011 - 12:10 AM

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.
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#4 fysio lab

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Posted 24 June 2011 - 12:38 AM

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

#5 lical208

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Posted 24 June 2011 - 01:45 AM

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?

Edited by lical208, 24 June 2011 - 03:18 AM.


#6 lical208

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Posted 24 June 2011 - 01:46 AM

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?

#7 bob1

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Posted 25 June 2011 - 05:35 PM

Yeah, mitochondria is a good option. You might want to try endoplasmic reticulum too.

#8 rhombus

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Posted 30 June 2011 - 04:38 AM

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.




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