Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa (ATCC® CCL-2™))
Permeabilization
Yes - 0.1% Triton X100 in PBS or 20UuM Digitonin in PBSpH 7.4
Specification
HeLa (ATCC® CCL-2™)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
Other product details
Incubation time
1 hour(s) and 30 minute(s) · Diluent: PBS pH 7.4
Dilution
1/250
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Goat anti Rabbit Alexa 555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Additional data
Additional Notes
HeLa cells were fixed for 15 min in 4%PFA, washed twice in PBS and subsequently permeabilized with either ,0.1% Triton X100, for 10 min at RT, or 20µm digitonin for 10 min at RT. Cells were blocked for 60 minutes in 10% normal goat serum (in PBS). ab2900 was diluted 1/250 in PBS and the antibody incubated 1h 30 min in a humidified chamber. Cells were extensively washed with PBS and incubated with goat anti-rabbit Alexa 555 (1/500) for 1h 30 min at RT. Finally, cells were mounted in Dako+ Hoechst 33342 and analyzed under a spinning disc confocal microscope.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Mauricio Terebiznik
Verified customer
投稿 Dec 21 2020