what cell line to use for visualizing membrane localized protein - (Oct/08/2008 )
Dear all,
I work with a cytosolic protein.
to this protein i attached a peptide that should induce plasma membrane localization.
i transfect this construct (peptide-bound) into HEK293 cells.
after staining with specific Ab i check for the localization to the plasma membrane using confocal microscope.
I can not clearly see that my protein localizes at the plasma membrane.
i would like to know whether HEK cells are ok for studying membrane localized proteins. If not what other cells should i use?
And is there any marker i can co stain to better see the plasma membrane?
Thank you in advance for support!
sarah
-sarah luke-
QUOTE (sarah luke @ Oct 8 2008, 09:35 AM)
Dear all,
I work with a cytosolic protein.
to this protein i attached a peptide that should induce plasma membrane localization.
i transfect this construct (peptide-bound) into HEK293 cells.
after staining with specific Ab i check for the localization to the plasma membrane using confocal microscope.
I can not clearly see that my protein localizes at the plasma membrane.
i would like to know whether HEK cells are ok for studying membrane localized proteins. If not what other cells should i use?
And is there any marker i can co stain to better see the plasma membrane?
Thank you in advance for support!
sarah
I work with a cytosolic protein.
to this protein i attached a peptide that should induce plasma membrane localization.
i transfect this construct (peptide-bound) into HEK293 cells.
after staining with specific Ab i check for the localization to the plasma membrane using confocal microscope.
I can not clearly see that my protein localizes at the plasma membrane.
i would like to know whether HEK cells are ok for studying membrane localized proteins. If not what other cells should i use?
And is there any marker i can co stain to better see the plasma membrane?
Thank you in advance for support!
sarah
here is a luseful ink to the Invitrogen Handbook:
http://probes.invitrogen.com/handbook/sections/1404.html
if your protein is not to detect at the membrane, where is it? peri nuclear pools? or still cytosolic?
-The Bearer-