Anti-PD-L2 PDCD1LG2 Antibody

Western Blot Validation in Human Raji Cell Lysate
Loading: 15 μg of lysates per lane. Antibodies: PD-L2 A03295 (A: 0.5 μg/mL and B: 1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines
Loading: 15 μg of lysates per lane. Antibodies: PD-L2, A03295 (4 μg/mL), competitor antibody (4 μg/mL), and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

KO Validation in HeLa Cells
Loading: 15 μg of HeLa WT cell lysates or PD-L2 KO cell lysates. Antibodies: PD-L2, A03295 (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

KO Validation in HeLa Cells
Loading: 15 μg of HeLa WT cell lysates or PD-L2 KO cell lysates. Antibodies: PD-L2, A03295 (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry Validation of PD-L2 in Mouse Brain Tissue
Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PD-L2 antibody (A03295) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of PD-L2 in Mouse Brain Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain cells labeling PD-L2 with A03295 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunohistochemistry Validation of PD-L2 in Mouse Brain Tissue
Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PD-L2 antibody (A03295) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of PD-L2 in Mouse Brain Tissue
Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain tissue labeling PD-L2 with A03295 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Regulated Expression Validation of PD-L2 in Mice with Melanoma Tumor (Knox et al., 2019)
Immunoblot analysis of PD-L2 expression with anti-PD-L2 (A03295) antibodies. PD-L2 expression was up-regulated by anti-PD1 antibody treatment whereas it was reduced by Next A alone or combination treatment (anti-PD1 antibody + NextA).

Immunohistochemistry Validation of PD-L2 in Lung Tumor of Mice (Kao et al., 2015)
Protein analysis for PD-L2 (E-H) by immunohistochemistry with anti-PD-L2 antibodies in mice lung tumors. hMUC1.Tg mice were induced with lung adenoma and then treated with concurrent or sequential cisplatin/radiotherapy. PD-L2 expression level at week 41 after treatment was similar in control and treatment groups.