CD300f/LMIR3/CD300LF Antibody (234903) [Unconjugated]

CD300f/LMIR3 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Rat Anti-Human CD300f/LMIR3 Monoclonal Antibody (Catalog # MAB2774) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready, ICC/IF
• Clone: 234903
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: LYOPH

CD300f/LMIR3/CD300LF Antibody (234903) [Unconjugated] Summary

• Immunogen: Y3 rat myeloid cell line transfected with human CD300f/LMIR3
• Specificity: Detects human CD300f/LMIR3 in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human LMIR1, 2, 4, 5, 6, or recombinant mouse LMIR3 is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Gene: CD300LF
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.25 ug/10^6 cells
  • CyTOF-ready
  • Immunocytochemistry 8-25 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 as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 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 CD300f/LMIR3/CD300LF Antibody (234903) [Unconjugated]

• CMRF35-like molecule 1
• CD300 molecule-like family member f
• CD300f
• CD300LF
• CLIM1
• CLM1
• CLM-1
• DIgR2
• IGSF13
• IREM1
• IREM-1
• LMIR3
• NKIR
• Pigr3

Background

CD300f, also known as CD300LF, LMIR3, IREM-1, CLM-1, IgSF13, DIgR1, and MAIR-V, is a 50‑60 kDa glycoprotein member of the immunoglobulin superfamily (1). Human CD300f consists of a 137 amino acid (aa) extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane segment, and a 113 aa cytoplasmic domain that contains multiple immunoreceptor tyrosine-based inhibitory motifs (ITIMs) or ITIM-like sequences (2, 3). Alternate splicing generates additional isoforms that carry substituted C-terminal tails of varying lengths and sequences following the ECD or transmembrane segment (3). Within the ECD, human CD300f shares 43% aa sequence identity with mouse and rat CD300f. CD300f is expressed on the surface of dendritic cells, monocytes, granulocytes, and mast cells as well as on acute myeloid leukemia (AML) blasts (2‑4). Pervanadate treatment or antibody crosslinking of CD300f induces phosphorylation of tyrosine residues in the cytoplasmic domain and the subsequent recruitment of phosphatases SHIP, SHP-1, SHP-2, and the p85 alpha subunit of PI3K (2, 3, 5, 6). CD300f ligation can induce cell death and inhibit signaling through multiple receptors including Fc epsilon RI, LMIR4, SCF R, TLR2, TLR3, and TLR9 (3‑8). In contrast, it enhances TLR4-mediated signaling/cytokine production in mast cells through association with the activating signaling protein FcR gamma (5). In mouse, a splice variant of CD300f (known as DIgR2, with a 7 aa insertion in the ECD) inhibits CD4+ T cell activation and in vivo Th1 and CTL responses (9). CD300f is upregulated on monocytes surrounding experimentally-induced spinal cord demyelination and functions as a negative regulator of inflammation in the CNS (10).

1. Clark, G.J. et al. (2009) Trends Immunol. 30:209.
2. Sui, L. et al. (2004) Biochem. Biophys. Res. Commun. 319:920.
3. Alvarez-Errico, D. et al. (2004) Eur. J. Immunol. 34:3690.
4. Korver, W. et al. (2009) Leukemia 23:1587.
5. Izawa, K. et al. (2009) J. Immunol. 183:925.
6. Alvarez-Errico, D. et al. (2007) J. Immunol. 178:808.
7. Can, I. et al. (2008) J. Immunol. 180:207.
8. Izawa, K. et al. (2007) J. Biol. Chem. 282:17997.
9. Shi, L. et al. (2006) Blood 108:2678.
10. Xi, H. et al. (2010) J. Exp. Med. 207:7.

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 CD300f/LMIR3/CD300LF Antibody (MAB2774)(2)

Publications using MAB2774

• Izawa K, Isobe M, Matsukawa T, Ito S, Maehara A, Takahashi M, Yamanishi Y, Kaitani A, Oki T, Okumura K, Kitamura T, Kitaura J Sphingomyelin and ceramide are physiological ligands for human LMIR3/CD300f, inhibiting FcepsilonRI-mediated mast cell activation. J Allergy Clin Immunol, 2014;133(1):270-3.e1-7. 2014 [PMID: 24035150] (Western Blot, Mouse)
• Asai, Tsuneaki, Morrison, Sherie L The SRC family tyrosine kinase HCK and the ETS family transcription factors SPIB and EHF regulate transcytosis across a human follicle-associated epithelium model. J Biol Chem, 2013;288(15):10395-405. 2013 [PMID: 23439650] (Neutralization, Human)

Bioinformatics

• Gene Symbol: CD300LF