Application
Flow Cytometry
Sample
Human Cell (PBMC)
Permeabilization
Yes - 4x Cytofix, 45 min, 4 ¯C
Gating Strategy
CD19 positive live cells
Specification
PBMC
Preparation
Cell harvesting/tissue preparation method: PBMC from whole blood by density gradient (lymphoprep, Fresenius)
Sample buffer: D-PBS+5 % FCS + 2 mM EDTA
Sample buffer: D-PBS+5 % FCS + 2 mM EDTA
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: D-PBS+5 % FCS + 2 mM EDTA
Dilution
1/100
Secondary antibody
Dilution
1/100
Name
Non-Abcam antibody was used: Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondar
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 488
Additional data
Additional Notes
We tried to stain permeablizied PBMCs, too. But it didn`t work as well as extracellular staining. Additional controls/antibody concentrations/incubation times have to be established if you want to stain permeabilized cells.
D-PBS+5 % FCS + 2 mM EDTA was replaced by PermWash (BD bioscience) when staining permeabilized cells.
Live/dead staining: Live/dead violett (1:1000 in PBS), 30 min, 4 ¯C
Blocking: FcR Blocking reagent human, Milteny Biotec, 10 min RT
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.
Clara Lettl
Verified customer
投稿 Jan 18 2017