This antbibody works nicely for immunostaining of lysosomes in HEK293 and HepG2 cell.
In my experience, permeabilization with Triton X-100 (0.1 %) works fine.
Antibody dilution: 1:200
NB! Note that this antibody stops working if stored at 4C. Should be stored in small aliquots at -20C.
Western blot
Not Rated
Immunostaining
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On February 15, 2012Simon Glerup, Aarhus wrote:
4
We use this antibody as a lysosomal marker in human-derived cell lines, e.g. 293 cells.
Dilution 1:200.
The image above shows human GDNF (green) bound to the surface of HEK293 cells transfected with the GDNF receptor GFRa1. Cells are also stained for Lamp-1 (red) using mAb H4A3.
NB. For mouse cell lines, we use Abcam 24170.
It is important to consider what detergent is used for permeabilization of cells. Saponin appears to give much better lysosome stainings than Triton X-100.
For inhibition of lysosomal degradation and better visualization of lysosomes, incubate with 50 ug/ml leupeptin and pepstatin for 24 h before fixing the cells. During this period, the leupeptin/pepstatin medium has to be changed every six hours.
On February 6, 2018 Julie Torvund-Jensen wrote:
This antbibody works nicely for immunostaining of lysosomes in HEK293 and HepG2 cell.
In my experience, permeabilization with Triton X-100 (0.1 %) works fine.
Antibody dilution: 1:200
NB! Note that this antibody stops working if stored at 4C. Should be stored in small aliquots at -20C.
On February 15, 2012 Simon Glerup, Aarhus wrote:
We use this antibody as a lysosomal marker in human-derived cell lines, e.g. 293 cells.
Dilution 1:200.
The image above shows human GDNF (green) bound to the surface of HEK293 cells transfected with the GDNF receptor GFRa1. Cells are also stained for Lamp-1 (red) using mAb H4A3.
NB. For mouse cell lines, we use Abcam 24170.
It is important to consider what detergent is used for permeabilization of cells. Saponin appears to give much better lysosome stainings than Triton X-100.
For inhibition of lysosomal degradation and better visualization of lysosomes, incubate with 50 ug/ml leupeptin and pepstatin for 24 h before fixing the cells. During this period, the leupeptin/pepstatin medium has to be changed every six hours.