Table 1.
Antibody Resource List
| Protein target | Staining type | Antibody step | Antigen | Source (catalog number) | Working concentration |
|---|---|---|---|---|---|
| Acta2 | IHC | Primary | FITC mouse anti-human Acta2 IgG2a | Millipore-Sigma, St. Louis, MO (F3777) | 2 μg/mL |
| Isotype | FITC mouse IgG2a | BD Biosciences, San Jose, CA (553456) | 2 μg/mL | ||
| Secondary | Biotinylated goat anti-FITC IgG | Vector Laboratories, Burlingame, CA (B0601) | 1 μg/mL | ||
| Vcam1 | IHC | Primary | Biotinylated rat anti-mouse Vcam1 IgG2a | BD Biosciences (553331) | 5 μg/mL |
| Isotype | Biotinylated rat IgG2a | Biolegend, San Diego, CA (400504) | 5 μg/mL | ||
| Secondary | ABC-Elite | Vector Laboratories (PK-6100) | n/a | ||
| Fap | Multiplex panel 1 | Primary | Biotinylated sheep anti-human Fap IgG | R&D Systems, Minneapolis, MN (BAF3715) | 5 μg/mL |
| Isotype | Biotinylated sheep IgG | R&D Systems (BAF020) | 5 μg/mL | ||
| Secondary | HRP donkey anti-sheep IgG | Jackson ImmunoResearch Laboratories, West Grove, PA (713-035-147) | 5 μg/mL | ||
| Tertiary | AF594-tyramide | Thermo Fisher Scientific, Waltham, MA (T20935) | 1:100 | ||
| Vcam1 | Multiplex panel 1 | Primary | Biotinylated rat anti-mouse Vcam1 IgG2a | BD Biosciences (553331) | 5 μg/mL |
| Isotype | Biotinylated rat IgG2a | Biolegend (400504) | 5 μg/mL | ||
| Secondary | BV711-streptavidin | Biolegend (405241) | 2 μg/mL | ||
| Vim | Multiplex panel 1 | Primary | Rabbit anti-human Vim IgG | Cell Signaling Technology, Danvers, MA (5741) | 1 μg/mL |
| Isotype | Rabbit IgG | Jackson ImmunoResearch Laboratories (011-000-003) | 1 μg/mL | ||
| Secondary | AF350 donkey anti-rabbit IgG | Thermo Fisher Scientific (A10039) | 5 μg/mL | ||
| Acta2 | Multiplex panel 1 | Primary | FITC mouse anti-human Acta2 IgG2a | Millipore-Sigma (F3777) | 5 μg/mL |
| Isotype | FITC mouse IgG2a | BD Biosciences (553456) | 5 μg/mL | ||
| Cd68 | Multiplex panel 1 | Primary | BV605 rat anti-mouse Cd68 IgG2a | Biolegend (137021) | 2 μg/mL |
| Isotype | BV605 rat IgG2a | Biolegend (400539) | 2 μg/mL | ||
| DRAQ5 | Multiplex panel 1 | Counterstain | NA | Biolegend (424101) | 1:1000 |
| Col1 | Multiplex panel 2 | Primary | Goat anti-human Col1 IgG | Southern Biotech, Birmingham, AL (1310-01) | 1 μg/mL |
| Isotype | Goat IgG | Jackson ImmunoResearch Laboratories (005-000-003) | 1 μg/mL | ||
| Secondary | AF488 donkey anti-goat IgG | Thermo Fisher Scientific (A11055) | 5 μg/mL | ||
| Fn1 | Multiplex panel 2 | Primary | Rabbit anti-human Fn1 IgG | Millipore-Sigma (F3648) | 1 μg/mL |
| Isotype | Rabbit IgG | Jackson ImmunoResearch Laboratories (011-000-003) | 1 μg/mL | ||
| Secondary | AF647 donkey ant-rabbit IgG | Thermo Fisher Scientific (A31573) | 5 μg/mL | ||
| Lama1 | Multiplex panel 2 | Primary | Rat anti-mouse Lama1 IgG2a | R&D Systems (MAB4656) | 1 μg/mL |
| Isotype | Rat IgG2a | eBioscience, San Diego, CA (16-4321) | 1 μg/mL | ||
| Secondary | AF594 donkey anti-rat IgG | Thermo Fisher Scientific (A21209) | 5 μg/mL | ||
| DAPI | Multiplex panel 2 | Counterstain | NA | Millipore-Sigma (10236276001) | 0.25 μg/mL |
| Cd68 | Multiplex panel 3 | Primary | Rat anti-mouse Cd68 IgG2a | Abcam, Cambridge, UK (53444) | 1 μg/mL |
| Isotype | Rat IgG2a | eBioscience (16-4321) | 1 μg/mL | ||
| Secondary | AF568 goat anti-rat IgG | Thermo Fisher Scientific (A11077) | 2 μg/mL | ||
| Nos2 | Multiplex panel 3 | Primary | Rabbit anti-mouse Nos2 IgG | Abcam (15323) | 1 μg/mL |
| Isotype | Rabbit IgG | Jackson ImmunoResearch Laboratories (011-000-003) | 1 μg/mL | ||
| Secondary | AF488 goat anti-rabbit IgG | Thermo Fisher Scientific (A11034) | 2 μg/mL | ||
| DAPI | Multiplex panel 3 | Counterstain | NA | Millipore-Sigma (10236276001) | 0.25 μg/mL |
| Cd68 | Multiplex panel 4 | Primary | Rat anti-mouse Cd68 IgG2a | Abcam (53444) | 1 μg/mL |
| Isotype | Rat IgG2a | eBioscience (16-4321) | 1 μg/mL | ||
| Secondary | AF568 goat anti-rat IgG | Thermo Fisher Scientific (A11077) | 2 μg/mL | ||
| Cd206 | Multiplex panel 4 | Primary | Rabbit anti-human Cd206 IgG | Abcam (64693) | 1 μg/mL |
| Isotype | Rabbit IgG | Jackson ImmunoResearch Laboratories (011-000-003) | 1 μg/mL | ||
| Secondary | AF488 goat anti-rabbit IgG | Thermo Fisher Scientific (A11034) | 2 μg/mL | ||
| DAPI | Multiplex panel 4 | Counterstain | NA | Millipore-Sigma (10236276001) | 0.25 μg/mL |
| Adgre1 | Multiplex panel 5 | Primary | Rat anti-mouse Adgre1 IgG2b | Bio-Rad, Hercules, CA (MCA497) | 4 μg/mL |
| Isotype | Rat IgG2b | Southern Biotech (0118-01) | 4 μg/mL | ||
| Secondary | AF488 donkey anti-rat IgG | Thermo Fisher Scientific (A21208) | 5 μg/mL | ||
| Cd68 | Multiplex panel 5 | Primary | BV605 rat anti-mouse Cd68 IgG2a | Biolegend (137021) | 2 μg/mL |
| Isotype | BV605 rat IgG2a | Biolegend (400539) | 2 μg/mL | ||
| Hmox1 | Multiplex panel 5 | Primary | Rabbit anti-human Hmox1 IgG | Abcam (68477) | 0.5 μg/mL |
| Isotype | Rabbit IgG | Jackson ImmunoResearch Laboratories (011-000-003) | 0.5 μg/mL | ||
| Secondary | AF647 donkey ant-rabbit IgG | Thermo Fisher Scientific (A31573) | 5 μg/mL | ||
| DAPI | Multiplex panel 5 | Counterstain | NA | Millipore-Sigma (10236276001) | 0.25 μg/mL |
Acta2, α-smooth muscle actin; Adgre1, adhesion G protein–coupled receptor E1; Col1, collagen type I; Fap, fibroblast activation protein; FITC, fluorescein isothiocyanate; Fn1, fibronectin; Hmox1, heme oxygenase; IHC, immunohistochemistry; Lama1, laminin; NA, not applicable; Nos2, nitric oxide synthase 2; Vcam1, vascular cell adhesion molecule 1; Vim, vimentin.