Recombinant Human LILRB2/CD85d/ILT4 Fc Avi-tag Protein, CF

When Recombinant Human Angiopoietin‑like Protein 2/ANGPTL2 C-Terminal Fragment (9795-AN) is immobilized at 1 µg/mL (100 µL/well), Biotinylated Recombinant Human LILRB2/CD85d/ILT4 Fc Chimera Avi-tag (Catalog # AVI2078) binds with an ED50 of 10-80 ng/mL.

2 μg/lane of Biotinylated Recombinant Human LILRB2/CD85d/ILT4 Fc Chimera Avi-tag Protein (Catalog # AVI2078) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 90-100 kDa and 180-200 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human LILRB2/CD85d/ILT4 Fc Avi-tag Protein, CF Summary

• Additional Information: Biotinylated
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human Angiopoietin-like Protein 2/ANGPTL2 C-Terminal Fragment (Catalog # 9795-AN) is immobilized at 1 µg/mL (100 µL/well), Biotinylated Recombinant Human LILRB2/CD85d/ILT4 Fc Chimera Avi-tag (Catalog # AVI2078) binds with an ED₅₀ of 10-80 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human LILRB2/CD85d/ILT4 protein
  

  Human ILT4 (Gln22-His458) Accession # NP_005865.3	IEGRMD	Human IgG1 Fc (Pro 100-Lys 330)	Avi-tag
  N-terminus			C-terminus

• Accession #: NP_005865.3
• N-terminal Sequence: No results obtained, Gln22 inferred from enzymatic pyroglutamate treatment revealing Thr23
• Structure / Form: Biotinylated via Avi-tag.
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 76 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 90-100 kDa, under reducing conditions.

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human LILRB2/CD85d/ILT4 Fc Avi-tag Protein, CF

• CD85 antigen-like family member D
• CD85d antigen
• CD85d
• Ig-like transcript 4
• ILT4
• ILT-4
• ILT4CD85d
• Immunoglobulin-like transcript 4
• Leukocyte immunoglobulin-like receptor 2
• leukocyte immunoglobulin-like receptor subfamily B member 2 soluble isoform
• leukocyte immunoglobulin-like receptor subfamily B member 2
• leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains)
• LILRB2
• LIR2
• LIR2CD85D
• LIR-2subfamily A (with TM domain), member 6
• member 2
• MIR10
• MIR10LILRA6
• Monocyte/macrophage immunoglobulin-like receptor 10

Background

The immunoglobulin-like transcript (ILT) comprise a family of activating and inhibitory type immunoreceptors whose genes are located in the same locus that encodes killer cell Ig-like receptors (KIR) (1-3). ILT4, also known as LIR-2 and LILRB2, is a type I transmembrane protein expressed primarily on monocytes and dendritic cells (DC) (4). Human ILT4 is produced as a 598 amino acid (aa) precursor including a 21 aa signal sequence, a 440 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 116 aa cytoplasmic domain. The ECD contains four Ig-like domains, and the cytoplasmic domain contains three immunoreceptor tyrosine-based inhibitory motifs (ITIM) (5). The ECD of human ILT4 shares 76% aa identity with chimpanzee ILT4 and 74%, 81%, 33%, 52%, 77%, 61%, and 64 % aa identity with human ILT1, 2, 3, 5, 6, 7, and 8, respectively. ILT4 binds to classical MHC I proteins as well as the non-classical HLA-G1 and HLA-F molecules (5-9). It competes with CD8 alpha for MHC I binding but does not compete with KIR2DL1 (7). Ligation of ILT4 induces Tyr phosphorylation within its cytoplasmic ITIMs, a requirement for association with SHP-1 (4, 6). Activation of ILT4 inhibits signaling through Fc gamma RI (4) and Fc epsilon RI (6) and causes DC to become tolerogenic by down-regulation of co‑stimulatory molecules (10, 11). ILT4 mediates tolerogenic DC‑induced CD4⁺ T cell energy in vitro and in vivo (10-12). Our Avi-tag Biotinylated human LILRB2/CD85d/ILT4 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

1. Suciu-Foca, N. et al. (2005) Int. Immunopharmacol. 5:7. 
2. Hofmeister, V. and E.H. Weiss (2003) Semin. Canc. Biol. 13:317. 
3. Hunt, J.S. et al. (2005) FASEB J. 19:681. 
4. Finger, N.A. et al. (1998) Eur. J. Immunol. 28:3423. 
5. Borges, L. et al. (1997) J. Immunol. 159:5192. 
6. Colonna, M. et al. (1998) J. Immunol. 160:3096. 
7. Shiroishi, M. et al. (2003) Proc. Natl. Acad. Sci. 100:8856.
8. Lepin, E.J.M. et al. (2000) Eur. J. Immunol. 30:3552.
9. Allen, R.L. et al. (2001) J. Immunol. 167:5543.
10. Chang, C.C. et al. (2002) Nat. Immunol. 3:237.
11. Ristich, V. et al. (2005) Eur. J. Immunol. 35:1133.
12. Manavalan, J.S. et al. (2003) Transpl. Immunol. 11:245.

Bioinformatics

• Uniprot:
  • NP_005865.3()