Ephrin B2 Antibody (YA3385)

Western blot analysis was performed on extracts from SH-SY5Y (lane 1, 15 μg), SK-OV-3 (lane 2, 15 μg), 293 (lane 3, 15 μg), Mouse brain (lane 4, 15 μg), and Rat brain (lane 5, 15 μg) using Ephrin B2 Rabbit mAb.Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:1000 dilution) and the loading control antibody (GAPDH, HY-P80137, 1:20000 dilution) were incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

Western blot analysis of extracts from SH-SY5Y(lane 2(20ug) and SH-SY5Y(lane 3(40ug) using Ephrin B2 Antibody (HY-P83640) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P83730, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

Immunocytochemistry analysis of C6 cells labeling Ephrin B2 with Ephrin B2 Antibody (HY-P83640) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with Ephrin B2 Antibody ((HY-P83640) at 1/50 dilution in BSA for Immunol Staining at 4 ℃,stay overnight. AF488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of HeLa cells labeling Ephrin B2 with Ephrin B2 Antibody (HY-P83640) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with Ephrin B2 Antibody ((HY-P83640) at 1/50 dilution in BSA for Immunol Staining at 4 ℃,stay overnight. AF488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).