Flow Cytometry
Detection of Treg (3-color) in Mouse Spleen
Source: Elabscience®Published: Dec 21,2023
|
Purpose |
Sample |
Antibody Collocation |
|
Adjust the voltage |
1 |
Blank |
|
Adjust compensation |
2 |
CD4-FITC |
|
3 |
CD25-APC |
|
|
4 |
Foxp3-PE |
|
|
APC-FMO in combination with Isotype Control for auxiliary gating |
5 |
CD4-FITC, Foxp3-PE; Rat IgG1, κ Isotype Control- APC |
|
PE-FMO in combination with Isotype Control for auxiliary gating |
6 |
CD4-FITC, CD25-APC; Mouse IgG1, κ Isotype Control-PE |
|
Full panel |
7 |
CD4-FITC, CD25-APC, Foxp3-PE |
|
Marker |
Fluorochrome |
Clone No. |
Cat. No. |
|
CD4 |
FITC |
GK1.5 |
|
|
CD25 |
APC |
PC-61.5.3 |
|
|
Foxp3 |
PE |
3G3 |
|
|
Rat IgG1, κ Isotype Control |
APC |
HRPN |
|
|
Mouse IgG1, κ Isotype Control |
PE |
MOPC-21 |
Tips:
1. Mouse Treg marker is CD4+ CD25+ Foxp3+.
2. CD4 cell population is obvious, and there is no need of Isotype Control. But CD25 and Foxp3 populations are not obvious, and Isotype Controls are needed.
3. There is fluorescence spillover, so it is necessary to set single positive tubes for compensation.
4. Please be careful that inappropriate use of Fixation/Permeabilization buffer may cause high background and unclear cell clustering.
