|
Purpose |
Sample |
Antibody Collocation |
|
|
Adjust the voltage |
1 |
Blank |
|
|
Adjust compensation |
2 |
CD3-FITC |
|
|
3 |
MHC II-Elab Fluor® Violet 450 |
||
|
4 |
CD11c-PE |
||
|
5 |
CD11b-APC |
||
|
Elab Fluor® Violet 450-FMO in combination with Isotype Control for auxiliary gating |
6 |
CD3-FITC、CD11c-PE、CD11b-APC、Rat IgG2b, κ Isotype Control-Elab Fluor® Violet 450 |
|
|
PE-FMO in combination with Isotype Control for auxiliary gating |
7 |
CD3-FITC、MHC II-Elab Fluor® Violet 450、CD11b-APC、Armenian Hamster IgG Isotype Control-PE |
|
|
APC-FMO in combination with Isotype Control for auxiliary gating |
8 |
CD3-FITC、MHC II-Elab Fluor® Violet 450、CD11c-PE、Rat IgG2b, κ Isotype Control-APC |
|
|
Full panel |
9 |
CD3-FITC、MHC II-Elab Fluor® Violet 450、CD11c-PE、CD11b-APC |
|
Marker |
Fluorochrome |
Clone No. |
Cat. No. |
|
CD3 |
FITC |
17A2 |
|
|
MHC II |
Elab Fluor® Violet 450 |
M5/114 |
|
|
CD11c |
PE |
N418 |
|
|
CD11b |
APC |
M1/70 |
|
|
Rat IgG2b, κ Isotype Control |
Elab Fluor® Violet 450 |
LTF-2 |
|
|
Armenian Hamster IgG Isotype Control |
PE |
PIP |
|
|
Rat IgG2b, κ Isotype Control |
APC |
LTF-2 |
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Tips:
1. Dendritic cells (DCs) in the mouse spleen are classified into lymphoid dendritic cells and myeloid dendritic cells.
2. The phenotype of lymphoid dendritic cells is CD11c⁺MHCII⁺CD11b⁻, while that of myeloid dendritic cells is CD11c⁺MHCII⁺CD11b⁺.
3. The staining protocol for the FMO (Fluorescence Minus One) control, used in conjunction with an isotype control, is identical to that of the test sample, except that the staining for one specific target marker is omitted and replaced with its corresponding isotype control. This FMO setup reveals the collective background fluorescence contributed by all other antibodies in the panel, thereby providing a critical reference for accurately establishing gating boundaries.
