Application
Flow Cytometry
Sample
Human Cell (HeLa)
Specification
HeLa
Fixation
Ice cold ethanol
Permeabilization
No
Gating Strategy
Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: PBS
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-Mouse Alexafluor 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/200
Additional data
Additional Notes
Asynchronous HeLa cells, were trypsinized, pelleted and fixed with ice cold ethanol overnight. 2x10^6 cells were immunofluorescently labeled for 1hr at room temp with the primary and then for 30 min with a secondary containing propidium iodide and RNase A at 37'C in the dark. This antibody gives a strong mitotic signal that was also confirmed by looking at the labeled cells under the microscope. This antibody does not give as strong a mitotic signal as the rabbit polyclonal ab5176, however it still clearly distinguishes the mitotic cells - there appears to be more background (i.e. interphase) staining in these cells which was confirmed by digital imaging microscopy.
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. Kirk Mcmanus
Verified customer
投稿 Nov 10 2008