Table 1.
Antibodies and counterstains used in this study
| Cells | Epitope | 1° Antibody, Clone, Source, Dilution | 2° Antibody, Source, Dilution |
|---|---|---|---|
| Endothelial Cells | CD31/Platelet-Endothelial Cell Adhesion Molecule | IF: Mouse anti-rat CD31 (Serotec) 1:100 FACS: Mouse anti-rat CD31(Chemicon) 1:100 | IF: Goat anti-mouse Cy3(Jackson ImmunoLabs) 1:1000; FACS: Goat anti- mouse PE (Caltag) 1:100 |
| Endothelial Cells/BM Stromal Cells | CD106/VCAM | IF & FACS: PE-conjugated Mouse anti- rat CD106 | IF: Goat anti-mouse Cy3(Jackson ImmunoLabs) 1:1000 |
| Mesenchymal Stem Cells | CD29/Beta-1 Integrin | IF& FACS: Biotin-conjugated CD29 (BD PharMingen) 1:100 | IF: Streptaviden Cy3(Jackson ImmunoLabs) 1:1000; FACS: Streptaviden PE (Cedarlane) 1:500 |
| Mesenchymal Stem Cells | CD90/Thy-1 | IF & FACS: PE-conjugated CD90 (BD PharMingen) 1:100 | IF: Streptaviden Cy3 (Jackson ImmunoLabs) 1:1000 |
| Endothelial Cell Precursors/Mesenchymal Stem Cells | CD133 | IF & FACS: Rabbit anti-CD133(GeneTex, Inc.) 1:100 | IF: Goat ant-rabbit Cy3(Jackson ImmunoLabs) 1:1000; FACS: Mouse ant- rabbit PE (SouthernBiotech) 1:100 |
| Leukocytes/Hematopoietic cells | CD45/Commo n Leukocyte Antigen | IF: Mouse anti-rat CD45, OX1 (Serotec) 1:100 FACS: Mouse anti-rat CD45 conjugated to PE, (BDPharMingen) 1:100 | IF: Goat anti-mouse Cy3 or Cy5 (Jackson ImmunoLabs) 1:1000 |
| Smooth Muscle Cells/pericytes | Smooth Muscle Actin | IF: Mouse anti-SMA conjugated to Cy3(Sigma) 1:250. | |
| GFP | Green Fluorescent Protein | TEM ImmunoGold: rabbit anti-GFP(Abcam) 1:100 | TEM ImmunoGold: Goat anti-rabbit 5 nm gold(Amersham) 1:25. |
| Counterstain | Nucleus | Hoechst 33258: Bis-benzimidizole(Sigma) 10 μg/ml | |
| Counterstain | F-actin | Phalloidin-Alexa 647 or Rhodamine- Phalloidin (Invitrogen) 1:250 |
IF: Immunofluorescence; FACS: Fluorescence activated cell sorting.
Vendor Source Addresses: Abcam: Cambridge, MA; Amersham: Piscataway, NJ; BD PharMingen, San Diego, CA; Caltag Laboratories: Burlingame, CA; Cedarlane: Ontario, Canada; Chemicon international, Temecula, CA; GeneTex: San Antonio, TX; Invitrogen, Carlesbad CA; Jackson ImmunoLabs, West Grove, PA; Serotec, Raleigh, NC; Sigma: St. Louis, MO; SouthernBiotech: Birmingham, AL