Anti psa

For 1D SDS-PAGE, protein samples were separated using reducing conditions on 12% polyacrylamide gels.^{11 (link)} Precision Plus Protein Standards (Bio-Rad, Hercules, CA, USA) were used to estimate apparent protein molecular mass. Immunoblot analysis was performed as previously described.⁹ Electroblots were incubated with the following primary antibody dilutions: anti-CD13 (1:5,000; Gene Tex Inc., Irvine, CA, USA.), anti-Mac-2-binding protein (M2BP) (1:1,000; R&D Systems, Minneapolis, MN, USA), anti-prostatic acid phosphatase (PAP) (1:50,000; Gene Tex Inc.), anti-PSA (1:5,000; Abcam, Cambridge, MA, USA), or anti-zinc α2-glycoprotein (ZAG) (1:50,000; Gene Tex Inc.).
Lectin blot analysis was performed as previously described,⁹ using biotinylated intact galectin-3 (10 µg/mL) or galectin-3 CRD (5.7 µg/mL). For competitive inhibition experiments, biotinylated galectin-3 was pre-incubated with 2 mg/mL asialofetuin, 200 mM lactose, or 200 mM sucrose in PBS-Tween 20 for two hours at 4 °C.
For 2D electroblot analysis of human seminal plasma, protein samples were separated in the first dimension using a Biorad-Protean IEF Cell isoelectric focusing apparatus. Isoelectrically focused proteins were subsequently run in the second dimension by SDS-PAGE to resolve proteins by molecular weight. Immunoblot and lectin analyses were performed a minimum of three times.