Application
Flow Cytometry
Fixation
None, cells were used fresh
Permeabilization
No
Sample
Mouse Cell (Cells from bronchoaveolar lavage in balb/c mice)
Specification
Cells from bronchoaveolar lavage in balb/c mice
Gating Strategy
alive (DAPI-),CD45+, C11b+, CD11c- cells
Preparation
Cell harvesting/tissue preparation method: Bronchoaveolar lavage in balb/c mice was performed with PBS-EDTA 3mM
Sample buffer: PBS-EDTA 3mM
Sample buffer: PBS-EDTA 3mM
Other product details
Incubation time
20 minute(s) · Temperature: 20°C · Diluent: PBS-ADTA 3mM
Dilution
1/2000
Secondary antibody
Name
Non-Abcam antibody was used: Jackson Immuno Research, anti-rat
Host species: Donkey
Clonality: Polyclonal
Conjugation: DyLight 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: DyLight 488
Dilution
1/1000
Additional data
Additional Notes
After incubation with antibody cocktail (CD45-APC, CD11b-PE, CD11c-AlexaFluor700 and NIMP-R14), cells were centrifuged (400g, 5min), rinsed with PBS-BSA 2%, centrifuged (400g, 5min) then secondary antibody and DAPI were added 20 min prior to count cells.
Black line represent isotype control and NIMP-R14 histogram is in pink. Sorted cells (ARIA, BD) confirm that NIMP-R14+ cells are Neutrophils.
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.
Mr. Francois Daubeuf
Verified customer
投稿 Feb 20 2015