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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 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
PD-L2 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. 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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