Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Human Cell (SHSY5Y)
Specification
SHSY5Y
Permeabilization
No
Fixative
Paraformaldehyde
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: PBS
Dilution
1/50
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti Rat AF488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
SHSY5Y cell line was grown on cover slips in 6 well plates in DMEM/10% FCS. Cells were washed with PBS prior to fixing with 1%PFA.
Fixed in 1%PFA prior to staining.
Wash 3x3min in PBS.
Block 10% goat serum for 20min at RT.
Primary Ab diluted in PBS and incubated overnight at 4deg.
Wash 3x3min in PBS.
Secondary – goat anti rat AF488 (green) 1/1000 in PBS for 1h at RT.
Wash 3x3min in PBS
Counterstained with Hoechst 33258 (blue) for 10mins at RT.
Wash 3min in dH2O
Coverslips removed tapped dry and placed on glass slides and mounted in Invitrogen ProLong gold and kept at 4deg until imaged.
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投稿 Apr 28 2014