Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Kelly Neuroblastoma cell line)
Permeabilization
No
Specification
Kelly Neuroblastoma cell line
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: PBS
Dilution
1/500
Secondary antibody
Dilution
1/1000
Name
Non-Abcam antibody was used: Invitrogen Goat Anti Chicken AF555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Additional data
Additional Notes
Kelly Neuroblastoma cell line was cultured in RPMI/10% FCS on coverslips in 12 well plates.
Cells were washed with PBS and fixed in 4% PFA until staining.
Staining protocol:
Wash 2x2min PBS
Serum block - 10% serum 20mins at RT
Primary - add primary in PBS for 2h at RT
Wash 2x2min PBS
Secondary – Goat anti Chicken AF555 (red) @ 1/1000 1h at RT
Wash 2x2min 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 Sep 26 2012