Application
Flow Cytometry
Fixation
Paraformaldehyde
Permeabilization
Yes - 1xPBS + 0.2% Triton X-100
Sample
Human Cell (Human osteosarcoma U2OS cells)
Specification
Human osteosarcoma U2OS cells
Gating Strategy
Single cells discriminated by DAPI staining
Preparation
Cell harvesting/tissue preparation method: Trypsinization
Sample buffer: 1xPBS + 1 mg/ml BSA + 1 microg/ml DAPI + 250 microg/ml RNase
Sample buffer: 1xPBS + 1 mg/ml BSA + 1 microg/ml DAPI + 250 microg/ml RNase
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: 1xPBS + 0.2% Triton X-100
Dilution
1/50
Secondary antibody
Name
Non-Abcam antibody was used: Anti-rabbit Molecular Probes
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
For this antibody to work well, permeabilisation has to be performed prior to fixation. The image shows proliferating cells on the left and cells treated with 1 microM camptothecin on the right. Camptothecin treatment results in RPA chromatinisation and enhanced signal in the S phase population.
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投稿 Aug 26 2014