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<strong>Figure 1 Western Blot Validation in Human Raji Cell Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: PD-L2 4063 (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.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: PD-L2, 4063 (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.
<strong>Figure 3 KO Validation in HeLa Cells</strong><br> Loading: 15 μg of HeLa WT cell lysates or PD-L2 KO cell lysates. Antibodies: PD-L2, 4063 (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.
<strong>Figure 3 KO Validation in HeLa Cells</strong><br> Loading: 15 μg of HeLa WT cell lysates or PD-L2 KO cell lysates. Antibodies: PD-L2, 4063 (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.
<strong>Figure 5 Immunohistochemistry Validation of PD-L2 in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PD-L2 antibody (4063) 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.
<strong>Figure 6 Immunofluorescence Validation of PD-L2 in Mouse Brain Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain cells labeling PD-L2 with 4063 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 7 Immunohistochemistry Validation of PD-L2 in Mouse Brain Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PD-L2 antibody (4063) 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.
<strong>Figure 8 Immunofluorescence Validation of PD-L2 in Mouse Brain Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse brain tissue labeling PD-L2 with 4063 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 9 Immunohistochemistry Validation of PD-L2 in Lung Tumor of Mice (Kao et al., 2015) </strong><br>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.
<strong>Figure 10 Regulated Expression Validation of PD-L2 in Mice with Melanoma Tumor (Knox et al., 2019) </strong><br>Immunoblot analysis of PD-L2 expression with anti-PD-L2 (4063) 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).
Figure 1 Western Blot Validation in Human Raji Cell Lysate
Loading: 15 μg of lysates per lane.Antibodies: PD-L2 4063 (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.
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PD-L2 Antibody

CATALOG NUMBER: 4063

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: rat (94%).
Immunogen:
Anti-PD-L2 antibody (4063) was raised against a peptide corresponding to 16 amino acids near the center of human PD-L2.
The immunogen is located within amino acids 140-190 of PD-L2.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 0.5-4 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in mouse samples; Immunofluorescence in mouse samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1207 - Raji Cell Lysate
Positive Control 2:
Cat. No. 1403 - Mouse Brain Tissue Lysate
Positive Control 3:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 4:
Cat. No. 1224 - Daudi Cell Lysate
Positive Control 5:
Cat. No. 1282 - 3T3/NIH Cell Lysate
Predicted Molecular Weight:
Predicted: 31kD
Observed: 29 kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar PD-L2 expression profile in human and mouse cell lines detected by two independent anti-PD-L2 antibodies that recognize different epitopes, 4063 against central domain and the competitor antibody against N-terminus domain.  PD-L2 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

KO Validation (Figure 3): Anti-PD-L2 antibodies (4063) specificity was further verified by PD-L2 specific knockout. PD-L2 signal was not detected in PD-L2 knockout HeLa cells as compared to that in control wild type cells.

Recombinant Protein Test (Figure 4): Anti-PD-L2 antibodies (4063) detected human PD-L2 recombinant protein at different concentrations.

Regulated expression validation (Figure 11): PD-L2 expression detected by anit-PD-L2 antibodies (4063) was up-regulated by anti-PD1 antibody treatment, which was reduced by Next A alone or combination treatment (anti-PD1 antibody + NextA).

Isoforms:
Human PD-L2 has 3 isoforms, including isoform 1 (273aa, 31kD), isoform 2 (183aa, 21kD) and isoform 3 (182aa, 21kD). Mouse PD-L2 has one isoform (247aa, 28kD) and Rat PD-L2 also has one isoform (268aa, 30kD). 4063 can detect human isoform and mouse isoform.
Purification:
PD-L2 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
Physical State:
Liquid
Buffer:
PD-L2 Antibody is supplied in PBS containing ≤0.09% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
PD-L2 antibody can be stored at 4°C, stable for up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
PDCD1LG2
Additional Names:
PD-L2 Antibody: B7DC, Btdc, PDL2, CD273, PD-L2, PDCD1L2, bA574F11.2, B7DC, Programmed cell death 1 ligand 2, Butyrophilin B7-DC, PD-1 ligand 2
Protein Accession Number:
NP_079515
Protein Gi Number:
190014605
Ncbi Gene Id Number:
80380
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
PD-L2 Antibody: Cell-mediated immune responses are initiated by T lymphocytes that are themselves stimulated by co gnate peptides bound to MHC molecules on antigen-presenting cells (APC). T-cell activation is generally self-limited as activated T cells express receptors such as PD-1 (also known as PDCD-1) that mediate inhibitory signals from the APC. PD-1 can bind two different but related ligands, PD-L1 and PD-L2, both of which are thought act as a negative regulator of T cell activation. However, it has been suggested that PD-L2 can act to stimulate an immunogenic response through and alternative receptor from PD-1.
Background Reference 1:
Holling et al. Hum. Immunol. 2004; 65:282-90.
Background Reference 2:
Ishida et al. EMBO J. 1992; 11:3887-95.
Background Reference 3:
LaGier and Pober. Hum. Immunol. 2006; 67:568-78.
Background Reference 4:
Zhang et al. Proc. Natl. Acad. Sci. USA 2006; 103:11695-700.
Citation 1:
Kao et al. Assessing the Effects of Concurrent versus Sequential Cisplatin/Radiotherapy on Immune Status in Lung Tumor-Bearing C57BL/6 Mice. Cancer Immunol Res. 2015;3(7):741-50. PMID: 25672395
Citation 2:
Knox et al. Selective HDAC6 inhibitors improve anti-PD-1 immune checkpoint blockade therapy by decreasing the anti-inflammatory phenotype of macrophages and down-regulation of immunosuppressive proteins in tumor cells. Sci Rep. 2019;9(1):6136. PMID: 30992475

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

4063

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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