Human OBCAM/OPCML Antibody AF2777: R&D Systems

Human OBCAM/OPCML Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human OBCAM/OPCML in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) KILON is observed and less than 1% cross-reactivity with rhLAMP is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human OBCAM/OPCML
    Gly28-Asn322
    Accession # Q14982
  • 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
    0.1 µg/mL
    Recombinant Human OBCAM (Catalog # 2777-CM)
  • Flow Cytometry
    2.5 µg/106 cells
    NG108‑15 mouse neuroblastoma/rat glioma hybrid cell line
  • Immunohistochemistry
    5-15 µg/mL
    Immersion fixed paraffin-embedded sections of human brain (cortex)
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
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
Immunocytochemistry
OBCAM/OPCML in MCF‑7 Human Cell Line. OBCAM/OPCML was detected in immersion fixed MCF‑7 human breast cancer cell line using Goat Anti-Human OBCAM/OPCML Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2777) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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: OBCAM/OPCML

OBCAM (Opioid-binding cell adhesion molecule, also known as OPCML) is a member of the IgLON family of cell adhesion molecules. All IgLONs are GPI-linked glycoproteins, contain three C2 type Ig-like domains, and are expressed in cerebral cortex and hippocampus (1, 2). The name IgLON derives from family membership in the Ig superfamily, and the first letters of the names of group’s molecules; LAMP, OBCAM, and Neurotrimin. Recently, membership in the group has been expanded by one with the addition of Kilon (Kindred of IgLON), and members of this group are now often referred to Diglons, based on the dimerizing nature of the IgLONs (1, 2). Human OBCAM is synthesized as a 345 amino acid (aa) preproprecursor that contains a 27 aa signal sequence, a 295 aa mature region, and a C‑terminal 23 aa prosegment (3). The prosegment is cleaved to generate the GPI-link. OBCAM varies in molecular weight, ranging from 46 kDa to 65 kDa (4‑6). The difference is not due to alternate splicing but to differential glycosylation (6). Although it is not unusual for GPI-linked proteins to be solubilized, to date there is no evidence that OBCAM functions as a soluble molecule (1). Mature human OBCAM is 98%, 99%, and 98% aa identical to mature bovine, rat and mouse OBCAM, respectively. OBCAM has limited expression, occurring principally in telencephalon and ovarian epithelium (7, 8). In brain, it is found associated with dendrites and post-synaptic membranes, where it may maintain synaptic architecture (1, 5). In ovary, it has been suggested to be a tumor-suppressor factor (8). The receptor(s) for OBCAM appears to be other members of the IgLON family, and a dimer is the functional unit. While neurotrimin appears to function as both a homodimer and heterodimer, all other family members (including OBCAM) show a preference for heterodimerization. OBCAM forms strong trans (between cells) heterodimers with LAMP, and modest heterodimers with Neurotrimin. There is hardly any binding with itself. Kilon likely binds OBCAM, but this interaction is not well studied (1). OBCAM heterodimers apparently bind to almost all possible IgLON heterodimer combinations on other cells. In cis (same cell), OBCAM also binds to LAMP and Neurotrimin (2).

  • References:
    1. Miyata, S. et al. (2000) Neuroscience 117:645.
    2. Reed, J. et al. (2004) J. Cell Sci. 117:3961.
    3. Shark, K.B. and N.M. Lee (1995) Gene 155:213.
    4. Wick, M.J. et al. (1996) Mol. Brain Res. 36:322.
    5. Miyata, S. (2000) J. Comp. Neurol. 242:74.
    6. Hachisuka, A. et al. (1996) Neurochem. Int. 28:373.
    7. Miyata, S. et al. (2003) Brain Res. 979:129.
    8. Sellar, G.C. et al. (2003) Nat. Genet. 34:337.
  • Long Name:
    Opioid-binding Cell Adhesion Molecule/Opioid Binding Protein/Cell Adhesion Molecule-like
  • Entrez Gene IDs:
    4978 (Human)
  • Alternate Names:
    IgLON family member 1; IGLON1opioid-binding protein/cell adhesion molecule-like; OBCAM; OBCAMopiate binding-cell adhesion molecule; OPCM; OPCML; opioid binding protein/cell adhesion molecule-like preprotein; opioid binding protein/cell adhesion molecule-like; Opioid-binding cell adhesion molecule; opioid-binding protein/cell adhesion molecule
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