Human/Mouse NCAM-1/CD56 Antibody

(2 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human NCAM-1/CD56 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross‑reactivity with recombinant human (rh) ALCAM, rhBCAM and rhEpCAM is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human NCAM1/CD56
    Leu20-Pro603
    Accession # NP_001070150
  • 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
    0.5 µg/mL
    See below
  • Simple Western
    5-25 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    Human peripheral blood mononuclear cells
  • 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
Detection of Human and Mouse
NCAM‑1/CD56 by Western Blot.
Western blot shows lysates of human brain (cerebellum and motor cortex) tissue and mouse brain (cerebellum) tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse NCAM‑1/CD56 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2408) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for NCAM‑1/CD56 at approximately 100-150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
NCAM-1/CD56 in SH-SY5Y Human Neuroblastoma Cells. SH-SY5Y human neuroblastoma cells were cultured overnight in the presence of 1 mM Retinoic Acid (Catalog # 0695/50) prior to immersion fixation. Neural Cell Adhesion Molecule 1 (NCAM-1)/CD56 was detected using a Goat Anti-Human/Mouse NCAM-1/CD56 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2408). The cells were stained with the NorthernLights 557-conjugated Donkey Anti-Goat IgG Affinity-purified Secondary Antibody (red; Catalog # NL001). Actin filaments were stained with FITC-conjugated Phalloidin (green) and cell nuclei were counter­stained with DAPI (blue). NCAM-1/CD56 immuno­reactivity was localized to the plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
NCAM‑1/CD56 in BG01V Human Embryonic Stem Cells. NCAM‑1/CD56 was detected in immersion fixed BG01V human embryonic stem cells differentiated into neural progenitor cells using Goat Anti-Human/Mouse NCAM‑1/CD56 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2408) 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Detection of Human and Mouse
NCAM‑1/CD56 by Simple WesternTM.
Simple Western lane view shows lysates of human and mouse brain (cerebellum) tissue, loaded at 0.2 mg/mL. A specific band was detected for NCAM‑1/CD56 at approximately 143 kDa (as indicated) using 5 µg/mL for human lysates and 25 µg/mL for mouse lysates of Goat Anti-Human/Mouse NCAM‑1/
CD56 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2408) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
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: NCAM-1/CD56

Neural cell adhesion molecule 1 (NCAM-1) is a multifunctional member of the Ig superfamily. It belongs to a family of membrane-bound glycoproteins that are involved in Ca++ independent cell matrix and homophilic or heterophilic cell-cell interactions. NCAM-1 specifically binds to heparan sulfate proteoglycans (1), the extracellular matrix protein agrin (2), and several chondroitin sulfate proteoglycans that include neurocan and phosphocan (3). There are three main forms of human NCAM-1 that arise by alternate splicing. These are designated NCAM-120/NCAM-1 (761 amino acids [aa]), NCAM-140 (848 aa), and NCAM-180 (1120 aa). NCAM-120 is GPI-linked, while NCAM-140 and NCAM-180 are type I transmembrane glycoproteins (4‑6). Additional alternate splicing adds considerable diversity to all three forms, and extracellular proteolytic processing is possible for NCAM-180 (7‑8). NCAM-1 is synthesized as a 761 aa preproprecursor that contains a 19 aa signal sequence, a 722 aa GPI-linked mature region, and a 20 aa C-terminal prosegment (4). The molecule contains five C-2 type Ig-like domains and two fibronectin type-III domains. Human to mouse, NCAM-1 is 93% aa identical. NCAM-1 appears to be highly sialylated. The polysialyation of NCAM-1 reduces its adhesive property and increases its neurite outgrowth promoting features (9). NCAM-1 in the adult brain shows a decline of sialylation relative to earlier developmental periods. In regions that retain a high degree of neuronal plasticity, however, the adult brain continues to express polysialylation-NCAM-1, suggesting sialylation of NCAM-1 is involved in regenerative processes and synaptic plasticity (10‑13).

  • References:
    1. Burg, M.A. et al. (1995) J. Neurosci. Res. 41:49.
    2. Storms, S.D. and U. Rutishauser (1998) J Biol. Chem. 273:27124.
    3. Margolis, R.K. et al. (1996) Perspect. Dev. Neurobiol. 3:273.
    4. Dickson, G. et al. (1987) Cell 50:1119.
    5. Lanier, L.L. et al. (1991) J. Immunol. 146:4421.
    6. Hemperly, J.J. et al. (1990) J. Mol. Neurosci. 2:71.
    7. Rutishauser, U.and C. Goridis (1986) Trends Genet. 2:72.
    8. Vawter, M.P. et al. (2001) Exp. Neurol. 172:29.
    9. Rutihauser, U. (1990) Adv. Exp. Med. Biol. 265:179.
    10. Becker, C.G. et al. (1996) J. Neurosci. Res. 45:143.
    11. Doherty, P. et al. (1995) J. Neurobiol. 26:437.
    12. Eckardt, M. et al. (2000) J. Neurosci. 20:5234.
    13. Muller, D. et al. (1996) Neuron 17:413.
  • Long Name:
    Neural Cell Adhesion Molecule
  • Entrez Gene IDs:
    4684 (Human); 17967 (Mouse); 24586 (Rat)
  • Alternate Names:
    CD56 / NCAM-1; CD56 antigen; CD56; MSK39; NCAM1; N-CAM-1; NCAM-1; NCAMantigen recognized by monoclonal 5.1H11; neural cell adhesion molecule 1; neural cell adhesion molecule, NCAM
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Modeling Developmental and Tumorigenic Aspects of Trilateral Retinoblastoma via Human Embryonic Stem Cells
    Authors: Y Avior, E Lezmi, D Yanuka, N Benvenisty
    Stem Cell Reports, 2017;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  2. Divergent modulation of neuronal differentiation by caspase-2 and -9.
    Authors: Pistritto G, Papaleo V, Sanchez P, Ceci C, Barbaccia ML
    PLoS ONE, 2012;7(5):e36002.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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Summary

ApplicationImmunohistochemistry
Sample TestedMouse brain cancer sample,Mouse brain cancer tissue
SpeciesMouse

Other Experimental Details

Other Experimental DetailsImmunohistochemistry-Floating section

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