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.
Monoclonal Rat IgG2A Clone # 232612
Protein A or G purified from hybridoma culture supernatant
Y3 rat myeloid cell line transfected with human CD300a/LMIR1
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.
Human granulocytes under non-reducing conditions only
2.5 µg/106 cells
Human whole blood
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.
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
Reconstitute at 0.5 mg/mL in sterile PBS.
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.
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).
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