Details and combinations of antibodies used for IF stainings
| Cell type . | Antibodies used for IF stainings . | ||||||
|---|---|---|---|---|---|---|---|
| Specificity . | Labeling . | *Sec. labeling . | Clone . | Isotype . | Dilution . | Manufacturer (location) . | |
| Leukocyte | CD45 | Purified | *A488 | 2D1 | Mouse IgG1 κ | 1:25 | BD Biosciences (Vienna, Austria) |
| T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter (Vienna, Austria) |
| CD4+/CD8+ | CD4 | Purified | *A568 | MT310 | Mouse IgG1 | 1:400 | Dako (Vienna, Austria) |
| CD8 | Purified | *A647 | C8/144B | Mouse IgG1 κ | 1:100 | Dako | |
| Regulatory T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD25 | FITC | *A568 | BC96 | Mouse IgG1 κ | 1:10 | BioLegend (California, CA) | |
| Foxp3 | Purified | *A647 | 236A/E7 | Mouse IgG1 | 1:300 | eBioscience (Vienna, Austria) | |
| NK T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD1d | Purified | *A568 | NOR3.2 | Mouse IgG1 | 1:50 | BioSource International (Camarillo, CA) | |
| NK cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD56 (Leu-19) | Purified | *A568 | MY31 | Mouse IgG1 κ | 1:100 | BD Biosciences | |
| B cell | CD19 | Purified | *A488 | J3-119 | Mouse IgG1 | 1:150 | Beckman Coulter |
| LC | CD1a | FITC | *A488 | HI149 | Mouse IgG1 κ | 1:50 | BD Pharmingen (Vienna, Austria) |
| CD207 (Langerin) | Purified | *A568 | DCGM4 | Mouse IgG1 | 1:1000 | Beckman Coulter | |
| pDC | CD303 (BDCA-2) | Purified | *A568 | 104C12.08 | Mouse IgG1 | 1:200 | Dendritics (Lyon, France) |
| CD123 (IL-3Ra) | Purified | *A488 | 9F5 | Mouse IgG1 | 1:100 | BD Pharmingen | |
| CD1c+ DC | CD1c (BDCA-1) | Purified | *A568 | M241 | Mouse IgG1 κ | 1:1000 | Ancell (Bayport, MN) |
| HLA-DR | APC | *A488 | G46-6 | Mouse IgG2a κ | 1:400 | BD Biosciences | |
| Macrophage | CD68 | Purified | *A568 | KP1 | Mouse IgG1 | 1:400 | Dako |
| CD163 | FITC | *A488 | 5C6-FAT | Mouse IgG1 | 1:100 | Acris (Herford, Germany) | |
| CD11c | Purified | *A647 | KB90 | Mouse IgG1 | 1:100 | Dako | |
| Neutrophil | CD15 | FITC | (no labeling) | 80H5 | Mouse IgM | 1:200 | Beckman Coulter |
| HLA-DR | APC | *A568 | G46-6 | Mouse IgG2a κ | 1:400 | BD Biosciences | |
| Eosinophil | MBP | Purified | *A488 | BMK13 | Mouse IgG1 | 1:400 | Research Diagnostics, Inc (Flanders, NJ) |
| Basophil | CD123 (IL-3Ra) | Purified | *A488 | 9F5 | Mouse IgG1 | 1:100 | BD Pharmingen |
| CD203c (ENPP-3) | Purified | *A568 | NP4D6 | Mouse IgG1 | 1:400 | BioLegend | |
| Mast cell | CD117 (c-Kit) | Purified | *A488 | 95C3 | Mouse IgG1 | 1:100 | An der Grub (Vienna, Austria) |
| CD203c (ENPP-3) | Purified | *A568 | NP4D6 | Mouse IgG1 | 1:400 | BioLegend | |
| Cell type . | Antibodies used for IF stainings . | ||||||
|---|---|---|---|---|---|---|---|
| Specificity . | Labeling . | *Sec. labeling . | Clone . | Isotype . | Dilution . | Manufacturer (location) . | |
| Leukocyte | CD45 | Purified | *A488 | 2D1 | Mouse IgG1 κ | 1:25 | BD Biosciences (Vienna, Austria) |
| T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter (Vienna, Austria) |
| CD4+/CD8+ | CD4 | Purified | *A568 | MT310 | Mouse IgG1 | 1:400 | Dako (Vienna, Austria) |
| CD8 | Purified | *A647 | C8/144B | Mouse IgG1 κ | 1:100 | Dako | |
| Regulatory T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD25 | FITC | *A568 | BC96 | Mouse IgG1 κ | 1:10 | BioLegend (California, CA) | |
| Foxp3 | Purified | *A647 | 236A/E7 | Mouse IgG1 | 1:300 | eBioscience (Vienna, Austria) | |
| NK T cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD1d | Purified | *A568 | NOR3.2 | Mouse IgG1 | 1:50 | BioSource International (Camarillo, CA) | |
| NK cell | CD3 | Purified | *A488 | UCHT1 | Mouse IgG1 κ | 1:150 | Beckman Coulter |
| CD56 (Leu-19) | Purified | *A568 | MY31 | Mouse IgG1 κ | 1:100 | BD Biosciences | |
| B cell | CD19 | Purified | *A488 | J3-119 | Mouse IgG1 | 1:150 | Beckman Coulter |
| LC | CD1a | FITC | *A488 | HI149 | Mouse IgG1 κ | 1:50 | BD Pharmingen (Vienna, Austria) |
| CD207 (Langerin) | Purified | *A568 | DCGM4 | Mouse IgG1 | 1:1000 | Beckman Coulter | |
| pDC | CD303 (BDCA-2) | Purified | *A568 | 104C12.08 | Mouse IgG1 | 1:200 | Dendritics (Lyon, France) |
| CD123 (IL-3Ra) | Purified | *A488 | 9F5 | Mouse IgG1 | 1:100 | BD Pharmingen | |
| CD1c+ DC | CD1c (BDCA-1) | Purified | *A568 | M241 | Mouse IgG1 κ | 1:1000 | Ancell (Bayport, MN) |
| HLA-DR | APC | *A488 | G46-6 | Mouse IgG2a κ | 1:400 | BD Biosciences | |
| Macrophage | CD68 | Purified | *A568 | KP1 | Mouse IgG1 | 1:400 | Dako |
| CD163 | FITC | *A488 | 5C6-FAT | Mouse IgG1 | 1:100 | Acris (Herford, Germany) | |
| CD11c | Purified | *A647 | KB90 | Mouse IgG1 | 1:100 | Dako | |
| Neutrophil | CD15 | FITC | (no labeling) | 80H5 | Mouse IgM | 1:200 | Beckman Coulter |
| HLA-DR | APC | *A568 | G46-6 | Mouse IgG2a κ | 1:400 | BD Biosciences | |
| Eosinophil | MBP | Purified | *A488 | BMK13 | Mouse IgG1 | 1:400 | Research Diagnostics, Inc (Flanders, NJ) |
| Basophil | CD123 (IL-3Ra) | Purified | *A488 | 9F5 | Mouse IgG1 | 1:100 | BD Pharmingen |
| CD203c (ENPP-3) | Purified | *A568 | NP4D6 | Mouse IgG1 | 1:400 | BioLegend | |
| Mast cell | CD117 (c-Kit) | Purified | *A488 | 95C3 | Mouse IgG1 | 1:100 | An der Grub (Vienna, Austria) |
| CD203c (ENPP-3) | Purified | *A568 | NP4D6 | Mouse IgG1 | 1:400 | BioLegend | |
Details of antibodies used for IF single/double/triple staining procedures and the antibody combinations used for the indicated cell types. Antibodies were, if indicated, labeled with the listed dyes (secondary labeling). *A488, antibody labeled with a Zenon Alexa Fluor 488 labeling kit; *A568, antibody labeled with Alexa Fluor 568; *A647, antibody labeled with Alexa Fluor 647); DC, dendritic cell; MBP, major basic protein; Sec., secondary.