Application
Flow Cytometry
Sample
Mouse Cell (Colonic entrochromaffin cells)
Permeabilization
No
Gating Strategy
EpCAM+CD45-Chga+
Specification
Colonic entrochromaffin cells
Preparation
Cell harvesting/tissue preparation method: colon dissected and mesentery removed. Feces flushed with PBS. Colon epithelium dissociated and enriched by moderate shaking in 2mM EDTA at 37°C for 15min. Supernatant recovered and washed twice in MACS buffer (PBS supplemented with 5% FBS and 2mM EDTA) for 10min, 300g. Pellet resuspended in 4ml of RPMI (supplemented with 2% FBS and Diapase II at final concentration of 1mg/ml and DNase at final concentration of 10ug/ml) and kept in 37°C water bath for 10min. Cells washed twice with MACS as before.
Sample buffer: PBS supplemented with 5% FBS and 2mM EDTA
Sample buffer: PBS supplemented with 5% FBS and 2mM EDTA
Fixation
Acetone
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS supp. with 5% FBS and 2mM EDTA
Dilution
1/200
Secondary antibody
Dilution
1/200
Name
Non-Abcam antibody was used: Jackson ImmunoResearch
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
Samples read on ImageStream
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投稿 May 29 2018