NCAM-1/CD56 Antibody [Unconjugated]

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.

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.

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.

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.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, Flow, CyTOF-ready, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

NCAM-1/CD56 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human NCAM1/CD56
  Leu20-Pro603
  Accession # NP_001070150
• 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.
• Details of Functionality: ActivityAssaywCitation not found None None None None
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: NCAM1

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Simple Western 5-25 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • CyTOF-ready
  • Immunocytochemistry 5-15 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 0.2 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 NCAM-1/CD56 Antibody [Unconjugated]

• 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

Background

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).

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.

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 NCAM-1/CD56 Antibody (AF2408)(2)

Publications using AF2408

• Y Avior, E Lezmi, D Yanuka, N Benvenisty Modeling Developmental and Tumorigenic Aspects of Trilateral Retinoblastoma via Human Embryonic Stem Cells Stem Cell Reports, 2017;0(0):. 2017 [PMID: 28392220] (ICC,, Human)
• Pistritto G, Papaleo V, Sanchez P, Ceci C, Barbaccia ML Divergent modulation of neuronal differentiation by caspase-2 and -9. PLoS ONE, 2012;7(5):e36002. 2012 [PMID: 22629307] (WB,, Human)

Bioinformatics

• Gene Symbol: NCAM1
• Entrez:
  • 4684(Human)
  • 17967(Mouse)
  • 24586(Rat)
• Uniprot:
  • NP_001070150()