Flow Cytometry
Detection of T/B/NK (4-color) in human peripheral blood
Source: Elabscience®Published: Dec 19,2023
|
Purpose |
Sample |
Antibody Collocation |
|
Adjust the voltage |
1 |
Blank |
|
Adjust compensation |
2 |
CD45-PerCP |
|
3 |
CD3-FITC |
|
|
4 |
CD16-PE/ CD56-PE |
|
|
5 |
CD19-APC |
|
|
PE-FMO in combination with Isotype Control for auxiliary gating |
6 |
CD45-PerCP, CD3-FITC, CD19-APC; Mouse IgG1, κ Isotype Control-PE |
|
Full panel |
7 |
CD45-PerCP, CD3-FITC, CD16-PE, CD56-PE, CD19-APC |
|
Marker |
Fluorochrome |
Clone No. |
Cat. No. |
|
CD45 |
PerCP |
HI30 |
|
|
CD3 |
FITC |
OKT3 |
|
|
CD16 |
PE |
3G8 |
|
|
CD56 |
PE |
5.1H11 |
|
|
CD19 |
APC |
CB19 |
|
|
Mouse IgG1, κ Isotype Control |
PE |
MOPC-21 |
Tips:
1. This panel has obvious cell populations, and compensation regulation can be performed without single positive tube. But for beginners of Flow Cytometry, it is recommended to set up a single positive tube to adjust compensation.
2. The detection standard for NK cells is CD3-CD16+CD56+.
3. In this panel, it is recommended to set Isotype Control for CD16 and CD56, while other indicators can be omitted due to obvious populations.
4. It is recommended to stain human peripheral blood samples with CD45, which is beneficial for the lymphocyte phylum gating through CD45 and SSC. It is recommended to use the single positive tube of CD45 to start the machine at low speed and set the threshold.
