Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (293 human embryonic kidney)
Specification
293 human embryonic kidney
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Other product details
Dilution
1/200
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 23°C · Diluent: 0,5%(w/v) saponin
Secondary antibody
Name
Non-Abcam antibody was used: goat anti rabbit AlexaFluor555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Dilution
1/500
Additional data
Additional Notes
Cells were plated on an uncoated cover slip.
One day after plating cells were washed twice with PBS and fixed with 4% PFA incl. 4% sucrose for 15 minutes at room temp.
Cell were permeabilized with 0.5% saponin in PBS for 45min at 4°C.
Blocking was carried out with 10% fetal calf serum in PBS-saponin during first and second antibody incubation.
After additional washing cover splips were mounted on glass slides with ProLong® Gold antifade reagent with DAPI (Invitrogen).
Antibody can probably be diluted further than 1:200.
One day after plating cells were washed twice with PBS and fixed with 4% PFA incl. 4% sucrose for 15 minutes at room temp.
Cell were permeabilized with 0.5% saponin in PBS for 45min at 4°C.
Blocking was carried out with 10% fetal calf serum in PBS-saponin during first and second antibody incubation.
After additional washing cover splips were mounted on glass slides with ProLong® Gold antifade reagent with DAPI (Invitrogen).
Antibody can probably be diluted further than 1:200.
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Submitted Sep 03 2010