Ab56301

Clinical specimens and xenografted lesions were subjected to immunohistochemistry (IHC) analysis as described previously [15 (link)]. The primary antibodies were anti-ANGPT2 (ab56301, Abcam) and anti-Annexin V antibody (ab14196, Abcam). The secondary antibodies were horseradish peroxidase-conjugated anti-rabbit immunoglobulin-G antibody (ab6721, Abcam).

The protein expressions of β-actin, VEGF, bFGF, Ang1, Ang2, PDGF-B, PDGF-D, and PlGF were studied using Western blotting. The primary antibodies used in this study were anti-β-actin (4970, Cell Signaling Technology, Danvers, MA, USA), anti-VEGF (sc-152, Santa Cruz Biotechnology, Dallas, TX, USA), anti-bFGF (sc-79, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-Ang1 (ab95230, Abcam, Cambridge, MA, USA), anti-Ang2 (ab56301, Abcam), anti-PDGF-B (ab23914, Abcam), anti-PDGF-D (40–2100, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), and anti-PlGF (sc-1883, Santa Cruz Biotechnology). After treatment with different hcy concentrations, the eyes were collected from the animals. A lysis buffer was used to extract retinal/choroidal proteins. The protein concentration was standardized, and the proteins were detected and quantified via electrochemiluminescence (Plus-ECL, PerkinElmer, Waltham, MA, USA) and densitometry.