TABLE 1.
Primary antibodies and controls. IB - Immunoblot, IHC - Immunohistochemistry, IF - Immunofluorescence, FACS - Flow cytometry, * from Dr. Derek Brown, UCB Cell Tech, Belgium.
| Antibody | Clone/Cat # | Supplier | Concentration used | Application |
|---|---|---|---|---|
| E-selectin | Mes-1 | Kind gift * | 10 μg/ml | IHC |
| VCAM-1 | CBL1300 | Merck Millipore | 10 μg/ml | IHC |
| ICAM-1 | YN1/1.7.4 | ATCC | 1:100 diluted supernatant | IHC |
| Phospho-NF-ĸB p65 (Ser536) | 3,033 | Cell signaling | 0.06 μg/ml | IB |
| NF-ĸB p65 | 8,242 | Cell signaling | 0.5 μg/ml | IB, IF |
| Phospho-p44/42MAPK (Thr202/Tyr204) | 9,101 | Cell signaling | 0.25 μg/ml | IB |
| p44/42 MAPK | 9,102 | Cell signaling | 0.003 μg/ml | IB |
| Axl | 4,566 | Cell signaling | 0.03 μg/ml | IB |
| Fyn | 4,023 | Cell signaling | 0.4 μg/ml, 10 μg/ml | IB, IHC, IF |
| Lck | 2,752 | Cell signaling | 0.2 μg/ml, 7.5 μg/ml | IB, IHC |
| GAPDH | sc-25778 | Santa Cruz Biotechnology | 0.04 μg/ml | IB |
| Axl | AF854 | RnDsystems | 2.5 μg/ml | IHC, IF |
| CD31 | 550,274 | BDPharmingen | 156 μg/ml | IF |
| APC CD106 | 305,810 | BioLegend | 3 µl in 100 µl staining volume | FACS |
| PE CD62E | 322,606 | BioLegend | 3 µl in 100 µl staining volume | FACS |
| FITC CD54 | 322,720 | BioLegend | 3 µl in 100 µl staining volume | FACS |
| Rat IgG1 | 0,116-01 | Southern Biotech | 10 μg/ml | IHC |
| Rat IgG2a | 0,117-01 | Southern Biotech | 10 μg/ml | IHC |
| Rat IgG2b | 0,118-01 | Southern Biotech | 10 μg/ml | IHC |
| Rabbit IgG | 0,111-01 | Southern Biotech | 10 μg/ml | IHC, IF |
| Goat IgG | 02-6,202 | Invitrogen | 2.5 μg/ml | IHC, IF |