Human PD-L2 / B7-DC Protein, mouse IgG1 Fc Tag

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
PDL2,PD-L2,Butyrophilin B7-DC,CD273,PDCD1 ligand 2,PDCD1L2,PDCD1LG2
Source
Human PD-L2, mouse IgG1 Fc Tag (PD2-H52A5) is expressed from human 293 cells (HEK293). It contains AA Leu 20 - Pro 219 (Accession # AAI13679).
Predicted N-terminus: Leu 20
Request for sequence
Molecular Characterization
This protein carreis a mouse IgG1 Fc tag at the C-terminus, and has a calculated MW of 48.9 kDa. The predicted N-terminus is Leu 20. The reducing (R) protein migrates as 60-70 kDa in SDS-PAGE due to glycosylation.
Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>95% as determined by reduced SDS-PAGE.

Formulation
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, pH7.5. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss is observed after storage at:
1. 4-8^°C for 12 months in lyophilized state;
2. -70^°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Human PD-L2, mouse IgG1 Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity

Immobilized Human PD-1, His Tag (Cat. No. PD1-H5221) at 2 μg/mL (100 μL/well) can bind Human PD-L2, Mouse IgG1 Fc Tag (Cat. No. PD2-H52A5) with a linear range of 10-156 ng/mL (QC tested).

Request for protocol

Human FcRn Heterodimer Protein Cell_Base

Flow Cytometry assay shows that recombinant Human PD-L2, mouse IgG1 Fc Tag (Cat.No. PD2-H52A5) can bind to 293 cell overexpressing human PD-1. The concentration of PD-L2 used is 0.1 ug/ml (Routinely tested).

Request for protocol

FACS analysis shows that the binding of Human PD-L2, mouse IgG1 Fc Tag to 293 overexpressing PD-1 was inhibited by increasing concentration of neutralizing anti-PD-1 antibody. The concentration of PD-L2 used is 0.1 ug/ml. The IC50 is 1.3 μg/mL (Routinely tested).

Request for protocol

Background
Programmed cell death 1 ligand 2 (PD-L2 or PDCD1 ligand 2) is also known as Butyrophilin B7-DC, CD antigen CD273, which belongs to the immunoglobulin superfamily or BTN/MOG family. The expression of PD-L2 is up-regulated by IFNG/IFN-gamma stimulation in monocytes and induced on dendritic cells grown from peripheral blood mononuclear cells with CSF2 and IL-4. PD-L2 Involved in the costimulatory signal, essential for T-cell proliferation and IFNG production in a PDCD1-independent manner. PD-L2 interaction with PDCD1 inhibits T-cell proliferation by blocking cell cycle progression and cytokine production.
References
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