ILT4/CD85d/LILRB2 Antibody [Unconjugated]

Recombinant Human LILRB2/CD85d/ILT4 Fc Chimera (Catalog # 2078-T4), immobilized onto a microplate, supports the adhesion of the HSB2 human peripheral blood acute lymphoblastic leukemia cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human LILRB2/CD85d/ILT4 Fc Chimera (35 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human LILRB2/CD85d/ILT4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2078). The ND50 is typically 0.3-1.0 µg/mL.

LILRB2/CD85d/ILT4 was detected in immersion fixed paraffin-embedded sections of Human Malignant Lymph Node using Goat Anti-Human LILRB2/CD85d/ILT4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2078) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Flow, IHC, B/N, CyTOF-ready
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

ILT4/CD85d/LILRB2 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human LILRB2/CD85d/ILT4
  Gly24-His458
  Accession # Q8N423
• Specificity: Detects human LILRB2/CD85d/ILT4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately
  35% cross-reactivity with recombinant human (rh) ILT2 is observed and 5% cross-reactivity with rhILT5 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: LILRB2
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunohistochemistry 3-25 ug/mL
  • Neutralization 0.3-1.0 ug/mL
  • Western Blot 0.1 ug/mL

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for ILT4/CD85d/LILRB2 Antibody [Unconjugated]

• 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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ILT4/CD85d/LILRB2 Antibody (AF2078)(3)

Publications using AF2078

• Yuki Kawaguchi, Junpei Matsubayashi, Yutaka Kawakami, Ryohei Nishida, Yuji Kurihara, Kohtaro Takei LOTUS suppresses amyloid beta -induced dendritic spine elimination through the blockade of amyloid beta binding to PirB Molecular Medicine 2022-12-12 [PMID: 36510132]
• M García, MB Palma, J Verine, S Miriuka, AM Inda, AL Errecalde, F Desgrandch, ED Carosella, D Tronik-Le The immune-checkpoint HLA-G/ILT4 is involved in the regulation of VEGF expression in clear cell renal cell carcinoma BMC Cancer, 2020-07-03;20(1):624. 2020-07-03 [PMID: 32620162] (IHC, Human)
• Gleissner CA, Zastrow A, Klingenberg R, Kluger MS, Konstandin M, Celik S, Haemmerling S, Shankar V, Giese T, Katus HA, Dengler TJ IL-10 inhibits endothelium-dependent T cell costimulation by up-regulation of ILT3/4 in human vascular endothelial cells. Eur. J. Immunol., 2007-01-01;37(1):177-92. 2007-01-01 [PMID: 17163451] (Neutralization, Human)

Bioinformatics

• Gene Symbol: LILRB2
• Uniprot:
  • Q8N423()