7 replies to this topic

[lical208]

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.

[zienpiggie]

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.

[Rsm]

lical208, on 23 June 2011 - 06:53 PM, 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.

[fysio lab]

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

[lical208]

fysio lab, on 24 June 2011 - 12:38 AM, 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?

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

[lical208]

fysio lab, on 24 June 2011 - 12:38 AM, 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?

[bob1]

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

[rhombus]

lical208, on 23 June 2011 - 06:53 PM, 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.