Recombinant Human PD-L2 Fc Biotinylated Protein, CF

Catalog #: BT1224 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT1224-050

Recombinant Human PD-L2 Fc Biotinylated Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human PD-L2 Fc Biotinylated Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human PD-1 Fc Chimera (Catalog # 1086-PD) is immobilized at 100 ng/mL, 100 μL/well, the concentration of Biotinylated Recombinant PD-L2 Fc Chimera that produces 50% of the optimal binding response is 0.04-0.24 μg/mL.

Source

Mouse myeloma cell line, NS0-derived human PD-L2/B7-DC protein

Human PD-L2 (Leu20-Pro219) Accession # Q9BQ51	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Biotinylated via sugars

Accession #

Q9BQ51

N-terminal Sequence
Analysis

Leu20

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

49 kDa (unlabeled)

Product Datasheets

BT1224

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

BT1224

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 250 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Recombinant Human PD-1 Fc Chimera Recom-binant Human PD-1 Fc Chimera (Catalog # 1086-PD) is coated at 0.1 µg/mL, Biotinylated Recom-binant Human PD-L2 Fc Chimera (Catalog # BT1224) binds with an ED₅₀ of 0.04-0.24 µg/mL.

Flow Cytometry

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In a functional flow cytometry test, (A) Recombinant Human PD-L2 Fc Biotinylated Protein (Catalog # BT1224) binds to HEK293 human embryonic kidney cell line transfected with recombinant human PD-1 and EGFP. Ligand binding was detected by staining cells with APC-conjugated Streptavidin (Catalog # F0050), which does not stain the cells in the absence of recombinant protein (B).

Reconstitution Calculator

Background: PD-L2/B7-DC

Programmed Death Ligand 2 (PD-L2), also known as B7-DC and butyrophilin-like protein, is a member of the B7 family of proteins that provide signals for regulating T-cell activation and tolerance (1). Mature human PD-L2 consists of a 201 amino acid (aa) extracellular domain (ECD) with one V-like and one C-like Ig domain, a 21 aa transmembrane segment, and a 32 aa cytoplasmic domain (2, 3). Within the ECD, mouse and human PD-L2 share 72% aa sequence identity. Alternative splicing generates additional isoforms that lack the second Ig-like domain and may be substituted and truncated following the first Ig-like domain (4). PD-L2 is expressed on dendritic cells, subsets of activated CD4⁺ and CD8⁺ T cells, and memory B cells that differentiate into plasma cells (3, 5, 6). At inflammatory sites such as rheumatoid arthritis, allergen exposure, and virus infection, PD-L2 is up-regulated on synoviocytes, infiltrating macrophages, dendritic cells, and airway epithelial cells (7-11). PD-L2, along with B7-H1/PD-L1, binds to T cell PD-1 where it promotes IFN-gamma production and CD40 Ligand up-regulation while inhibiting IL-4 production (2, 3, 12, 13). In addition, PD-L2 binds to RGM-B on macrophages and alveolar epithelial cells, supporting respiratory immune tolerance (14). In asthma, PD-L2 suppresses IL-5 and IL-13 production, promotes IL-12 production by dendritic cells, and supports allergen-induced airway hyper-responsiveness and mucus production (9, 11).

References

Ceeraz, S. et al. (2013) Trends Immunol. 34:556.
Latchman, Y. et al. (2001) Nat. Immunol. 2:261.
Tseng, S.-Y. et al. (2001) J. Exp. Med. 193:839.
He, X.-H. et al. (2004) Acta Biochim. Biophys. Sinica 36:284.
Messal, N. et al. (2011) Mol. Immunol. 48:2214.
Zuccarino-Catania, G.V. et al. (2014) Nat. Immunol. 15:631.
Guo, G. et al. (2012) Clin. Rheumatol. 31:271.
Loke, P. and J.P. Allison (2003) Proc. Natl. Acad. Sci. USA 100:5336.
Matsumoto, K. et al. (2004) J. Immunol. 172:2530.
Stanciu, L.A. et al. (2006) J. Infec. Dis. 193:404.
Lewkowich, I.P. et al. (2013) Mucosal Immun. 6:728.
Ghiotto, M. et al. (2010) Int. Immunol. 22:651.
Shin, T. et al. (2003) J. Exp. Med. 198:31.
Xiao, Y. et al. (2014) J. Exp. Med. 211:943.

Long Name

Programmed Death-1 Ligand-2

Entrez Gene IDs

80380 (Human); 58205 (Mouse); 309304 (Rat); 102146227 (Cynomolgus Monkey)

Alternate Names

B7-DC; bA574F11.2; Btdc; Butyrophilin B7-DC; Butyrophilin-like Protein; CD273 antigen; CD273; CD273PD-1 ligand 2; MGC142240; PD-1-ligand 2; PDCD1L2MGC142238; PDCD1LG2; PDL2; PD-L2; PDL2B7DC; PD-L2PDCD1 ligand 2; programmed cell death 1 ligand 2; Programmed death ligand 2