Table II.
Procedures for gal-3 and MDM2 staining.
| Protocol | Gal-3 | MDM2 |
|---|---|---|
| Blocking method | Horse seruma, 20 min, RT | Goat seruma, 20 min, RT |
| Primary antibody, dilution, incubation duration, incubation temperature, cat. no. | Anti-galectin-3, 1:1,000 in PBS, 16 h, 4°C; NCL-GAL3b | Anti-MDM2, 1:100 in PBS, 16 h, 4°C, NCL-MDM2b |
| Secondary antibody, dilution, incubation duration, incubation temperature, cat. no. | Biotynilated anti-mouse IgGa, 30 min, RT; PK-6100 | Biotinylated goat anti-mouse IgM, 30 min, RT ZMB2020c |
| Detection of secondary antibody | ABC-complexa, 30 min | ABC-complexa, 30 min |
| Chromogen | 1 mg/ml DABd, 5 min | 1 mg/ml DABd, 1 min |
a
Vectastain ABC kit; Vector Laboratories, Inc., Burlingame, CA, USA.
b
Novocastra; Leica Microsystems GmbH, Wetzlar, Germany.
c
Linaris GmbH, Dossenheim, Germany.
d
Dako, Glostrup, Denmark. Gal-3, galectin-3; MDM2, MDM2 proto-oncogene; Ig, immunoglobulin; DAB, 3,3′-diaminobenzidine; RT, room temperature.