Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SH-SY5Y)
Specification
SH-SY5Y
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
Other product details
Dilution
1/50
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS
Secondary antibody
Name
Non-Abcam antibody was used: Invitrogen Donkey Anti-Rabbit AF488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
SH-SY5Y neuroblastoma cell line differentiated using retinoic acid and BDNF, grown on glass cover slips in 6 well plates.
Fixed in 1%PFA prior to staining.
Wash 3x3min in PBS.
Block 10% donkey serum for 20min at RT.
Primary Ab diluted 1/50 in PBS and incubated overnight at 4deg, also works by incubating for 2 hours at RT.
Wash 3x3min in PBS.
Secondary - donkey anti-rabbit 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.
Fixed in 1%PFA prior to staining.
Wash 3x3min in PBS.
Block 10% donkey serum for 20min at RT.
Primary Ab diluted 1/50 in PBS and incubated overnight at 4deg, also works by incubating for 2 hours at RT.
Wash 3x3min in PBS.
Secondary - donkey anti-rabbit 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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提交于 Oct 07 2011