PD-L2 Mouse Monoclonal Antibody [A10D8]

☑ Relative expression (RE)

Western blot analysis of PD-L2 on different lysates with Mouse anti-PD-L2 antibody (HA601331) at 1/1,000 dilution.

Lane 1: Saos-2 cell lysate (hot lysis)
Lane 2: Jurkat cell lysate (negative)
Lane 3: MG-63 cell lysate (hot lysis)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 50 kDa

Exposure time: Lane 1-2: 1 minute; Lane 3: 2 minutes;
ECL: K1802;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601331) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

Immunocytochemistry analysis of HDLM-2 (positive) and Jurkat (negative) labeling PD-L2 with Mouse anti-PD-L2 antibody (HA601331) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PD-L2 antibody (HA601331) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.