Table 1.
Antibodies Used for Immunohistochemical Staining of Mouse Lungs
| Cells or molecules | Marker | Host | Manufacturer | Catalog no. | Comments |
|---|---|---|---|---|---|
| Lymphatic ECs | EGFP in Prox1-EGFP mice | Chicken | Aves (Davis, CA) | GFP 1020 | Overall best marker: stains nucleus and cytoplasm |
| Lymphatic ECs | Prox1 | Rabbit Goat |
AngioBio (Del Mar, CA) R&D Systems (Minneapolis, MN) |
11-002P AF2727 |
Stains lymphatic ECs, cardiac myocytes, and neuroendocrine cells |
| Lymphatic ECs | Vegfr3 | Goat | R&D Systems | AF743 | Weaker signal and worse background than Prox1 |
| Lymphatic ECs | Lyve1 | Rabbit Rat Goat |
AngioBio R&D Systems R&D Systems |
11-034 MAB2125 AF2125 |
Good in trachea, not in lungs; strongly expressed by some lung blood vessels |
| ECs | Pecam1 (Cd31) | Armenian hamster Rat |
Thermo Scientific (Rockford, IL) BioLegend (San Diego, CA) |
MA3105 102502 |
Weaker expression on lymphatic ECs than on blood vessels |
| ECs | Vegfr2 | Goat | R&D Systems | AF644 | Vascular ECs and more weakly on lymphatic ECs |
| High endothelial venules | MECA-79 | Rat (IgM) | BD Bioscience (San Jose, CA) | 553863 | In bronchoalveolar lymphoid tissue |
| Smooth muscle cells, myofibroblasts | α-Smooth muscle actin (Acta2) | Mouse | Sigma (St. Louis, MO) Sigma |
F3777 (FITC) C6198 (Cy3) |
Marks smooth muscle cells of large airways and blood vessels |
| Type 1 collagen | Type I collagen (Col1a1) | Rabbit Goat |
Millipore (Burlington, MA) Southern Biotech (Birmingham, AL) |
AB765P 1310-01 |
Used for most staining Colocalized with AB765P |
| Fibrin | Fibrin(ogen) | Rabbit | Dako (Carpinteria, CA) | A0080 | Recognizes both fibrin and fibrinogen |
| Platelets | Integrin a2b Itga2b, Cd41 |
Rat | BD BioSciences | 553847 | Clone MWReg30 |
| Leukocytes | Cd45 | Rat | BioLegend | 103102 | Clone 30-F11 |
| T lymphocytes | Cd3e | Armenian hamster |
BioLegend | 100302 | Clone 145-2C11 |
| B lymphocytes | B220 | Rat | BioLegend | 103202 | Clone RA3-6B2 |
| Neutrophils | S100a8 A100a9 |
Goat Rabbit |
R&D Systems Abcam (Burlingame, CA) |
AF3059 ab105472 | Also known as MRP8 and MRP14 |
| Macrophages | Aif1/Iba1 Cd11b Cx3cr1-EGFP Mrc1/Cd206 |
Rabbit Rat Chicken Rabbit |
WAKO (Richmond, VA); eBioscience (San Diego, CA) Aves Bio-Rad (Hercules, CA) |
019-19741 14-0112-82 GFP 1020 MCA2235GA |
Used for most staining Clone M1/70 Antibody to GFP Also stains alveolar macrophages and initial lymphatics |
| Apoptotic cells | Activated caspase-3 | Rabbit | R&D Systems | AF835 | |
| VEGF-C | Vegfc | Goat | Santa Cruz (Dallas, TX) | sc-7132 | No longer available |
Aif, allograft inflammatory factor; EC, endothelial cell; EGFP, enhanced green fluorescent protein; FITC, fluorescein isothiocyanate; GFP, green fluorescent protein; Iba, ionized calcium binding adapter molecule; Lyve, lymphatic vessel endothelial hyaluronan receptor; Pecam, platelet endothelial cell adhesion molecule; Prox, prospero homeobox; VEGF, vascular endothelial growth factor; Vegfr, VEGF receptor.