Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK 293T Transfected with SULF2 over expressing pl)
Permeabilization
Yes - .1% Triton x-100 in 1XPBS with 1 % Goat Serum
Specification
HEK 293T Transfected with SULF2 over expressing pl
Blocking step
5% Goat serum in 1XPBS with 0.01% Triton X-100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Other product details
Incubation time
22 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1XPBS 0.01% Triton
Dilution
1/500
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Invitrogen Goat anti-Mouse IgG, Alexa Fluor 488 (C
Host species: Goat
Clonality: Monoclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Monoclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
: ICC/IF using ab113405 to stain Sulfatase 2 in HEK293 cells transfected with a construct overexpressing human Sulfatase 2. Lipofectamine LTX (Invitrogen) was used for the transfection, in accordance with the manufacturer’s instructions. Two days post-transfection, cells were fixed with 4% (w/v) paraformaldehyde (15 min, 25°C), permeabilized in 1XPBS containing 0.1% triton X-100 and 1% goat serum (15 min, 25°C), and blocked with 1XPBS containing 5% goat serum and 0.01% triton X-100 (1hr, 25°C). Cells were incubated (22hr, 4°C) with ab113405 diluted 1/500 in 1XPBS containing 0.01% triton X-100 and subsequently blocked (1hr, 25°C) with 1XPBS containing 5% goat serum and 0.01% triton X-100. Finally, cells were incubated (1hr, 25°C) with Goat anti-Mouse Alexa Fluor 488 secondary antibody (Invitrogen) diluted 1/500 in 1XPBS containing 0.01% triton X-100. Nuclei were counterstained with DAPI. Control cells transfected with a construct overexpressing mCherry did not exhibit positive staining (not shown).
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投稿 Jul 30 2015