Flow Cytometry
Detection of Treg (6-color) in Human Peripheral Blood
Source: Elabscience®Published: Dec 21,2023
|
Purpose |
Sample |
Antibody Collocation |
|
Adjust the voltage |
1 |
Blank |
|
Adjust compensation |
2 |
CD45-Elab Fluor® Violet 450 |
|
3 |
CD3-Elab Fluor® Red 780 |
|
|
4 |
CD4-FITC |
|
|
5 |
CD8a-PerCP/Cyanine5.5 |
|
|
6 |
CD25-PE |
|
|
7 |
CD127-Elab Fluor®647 |
|
|
Full panel |
8 |
CD45-Elab Fluor® Violet 450, CD3-Elab Fluor® Red 780, CD4-FITC, CD8a-PerCP/Cyanine5.5, CD25-PE, CD127-Elab Fluor®647 |
|
Marker |
Fluorochrome |
Clone No. |
Cat. No. |
|
CD45 |
Elab Fluor® Violet 450 |
HI30 |
|
|
CD3 |
Elab Fluor® Red 780 |
OKT3 |
|
|
CD4 |
FITC |
RPA-T4 |
|
|
CD8a |
PerCP/Cyanine5.5 |
OKT-8 |
|
|
CD25 |
PE |
BC96 |
|
|
CD127 |
Elab Fluor®647 |
A019D5 |
Tips:
1. The traditional method of Foxp3 detection for Treg identification is relatively complicated, and the procedure of Fixation & Permeabilization requires high requirements for experimenters. Currently, CD127 is used for detection of human Treg.
2. Increase CD45 indicators to facilitate gating lymphocytes. The lymphocyte can be gated directly through CD45 and SSC, and analyze the cell ratio of CD4+CD25+CD127-/low, The proportion of Treg cells in lymphocytes of normal human peripheral blood ranges from 3% to 10% .
3. This scheme needs to set single positive tubes for compensation adjustment.
