Human CD300a/LMIR1 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human CD300a/LMIR1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, 100% cross‑reactivity with recombinant human LMIR2 (CMRF-35A) is observed and no cross-reactivity with recombinant human LMIR3, 4, 5, or 6 is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 232612
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Y3 rat myeloid cell line transfected with human CD300a/LMIR1
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    Human granulocytes under non-reducing conditions only
  • Flow Cytometry
    2.5 µg/106 cells
    Human whole blood
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human CD300a/LMIR1 by Western Blot. Western blot shows lysates of human granulocytes. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human CD300a/LMIR1 Monoclonal Antibody (Catalog # MAB2640) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for CD300a/LMIR1 at approximately 60 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD300a/LMIR1
CD300a, also known as LMIR1 (in rodents), CMRF-35H, IRp60, CLM-8, and MAIR-I, is a 60 kDa glycoprotein member of the immunoglobulin superfamily (1). Human CD300a consists of a 163 amino acid (aa) extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane segment, and a 98 aa cytoplasmic domain that contains three immunoreceptor tyrosine-based inhibitory motifs (ITIMs) and a non-canonical ITIM (2). Alternative splicing may generate additional isoforms that either lack the Ig-like domain or contain only the cytoplasmic domain. Within the ECD, human CD300a shares 40% and 43% aa sequence identity with mouse and rat LMIR1, respectively. In human, CD300a is expressed on peripheral blood eosinophils, mast cells, neutrophils, plasmacytoid dendritic cells, and various T cell subsets (3‑7). Antibody crosslinking of CD300a induces phosphorylation of tyrosine residues in the cytoplasmic domain. This leads to the recruitment of phosphatases SHIP, SHP-1, and SHP-2 and inhibition of NK cell, eosinophil, and mast cell activation (2, 3, 5‑7). Crosslinking of CD300a to other surface proteins such as SCF R or Fc epsilon RI on mast cells, Fc gamma RIIA on neutrophils, or CCR3 on mast cells and eosinophils inhibits downstream signaling from those receptors (5, 10‑12). CD300a crosslinking also limits the in vivo activities of these cells with a subsequent reduction of allergic inflammation symptoms (4, 11, 12).
  • References:
    1. Clark, G.J. et al. (2009) Trends Immunol. 30:209.
    2. Cantoni, C. et al. (1999) Eur. J. Immunol. 29:3148.
    3. Munitz, A. et al. (2006) Blood 107:1996.
    4. Bachelet, I. et al. (2005) J. Immunol. 175:7989.
    5. Alvarez, Y. et al. (2008) Mol. Immunol. 45:253.
    6. Ju, X. et al. (2008) Blood 112:1184.
    7. Clark, G.J. et al. (2007) J. Leukoc. Biol. 82:1126.
    8. Kumagai, H. et al. (2003) Biochem. Biophys. Res. Commun. 307:719.
    9. Yotsumoto, K. et al. (2003) J. Exp. Med. 198:223.
    10. Bachelet, I. et al. (2008) J. Immunol. 180:6064.
    11. Bachelet, I. et al. (2006) J. Allergy Clin. Immunol. 117:1314.
    12. Munitz, A. et al. (2006) J. Allergy Clin. Immunol. 118:1082.
  • Entrez Gene IDs:
    11314 (Human); 217303 (Mouse)
  • Alternate Names:
    CD300a antigenCMRF35H9; CD300a molecule; CD300a; CLM8; CLM-8; CMRF35H leukocyte immunoglobulin-like receptor; CMRF-35H; CMRF35-H; CMRF35-H9; CMRF-35-H9CD300 antigen-like family member A; CMRF35HIRp60; CMRF35-like molecule 8; IGSF12; IGSF12NK inhibitory receptor; Immunoglobulin superfamily member 12; Inhibitory receptor protein 60; IRC1; IRC1/IRC2; IRC2; IRp60; leukocyte membrane antigen; LMIR1; MAIR-I; Mcipr1; NKRL; Pigr4
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

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Rat IgG VisUCyte HRP Polymer Antibody

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Goat Anti-Rat IgG Antibody

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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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