Human/Mouse NCAM‑1/CD56 Antibody

Catalog # AF2408 | R&D Systems, Inc. a Bio-Techne Brand
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AF2408
AF2408-SP
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Key Product Details

Species Reactivity

Human, Mouse

Applications

CyTOF-ready, Flow Cytometry, Immunocytochemistry, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Summary for Human/Mouse NCAM‑1/CD56 Antibody

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse NCAM‑1/CD56 Antibody

Detection of Human and Mouse NCAM-1/CD56 antibody by Western Blot.

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.
NCAM-1/CD56 antibody in SH-SY5Y Human Neuroblastoma Cells by Immunocytochemistry (ICC).

NCAM-1/CD56 in SH-SY5Y Human Neuroblastoma Cells.

SH-SY5Y human neuroblastoma cells were cultured overnight in the presence of 1 μM Retinoic Acid (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 counter-stained with DAPI (blue). NCAM-1/CD56 immuno-reactivity was localized to the plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
NCAM-1/CD56 antibody in BG01V Human Embryonic Stem Cells by Immunocytochemistry (ICC).

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.
View 2 more images

Applications for Human/Mouse NCAM‑1/CD56 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed SH-SY5Y human neuroblastoma cell line and BG01V human embryonic stem cells differentiated into neural progenitor cells

Knockout Validated

NCAM‑1/CD56 is specifically detected in U937 human histiocytic lymphoma parental cell line but is not detectable in NCAM‑1/CD56 knockout U937 human histiocytic lymphoma cell line.

Simple Western

5-25 µg/mL
Sample: Human and mouse brain (cerebellum) tissue

Western Blot

0.5 µg/mL
Sample: Human brain (cerebellum and motor cortex) tissue and mouse brain (cerebellum) tissue
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using AF2408 in the following applications:

Formulation, Preparation and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in 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

Alternate Names

CD56, NCAM1

Entrez Gene IDs

4684 (Human); 17967 (Mouse); 24586 (Rat)

Gene Symbol

NCAM1

UniProt

NP_001070150

Product Documents for Human/Mouse NCAM‑1/CD56 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse NCAM‑1/CD56 Antibody

For research use only

Citations for Human/Mouse NCAM‑1/CD56 Antibody (13)

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Customer Reviews for Human/Mouse NCAM‑1/CD56 Antibody (2)

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  • Human/Mouse NCAM-1/CD56 Antibody
    Name: WonHee Kim
    Application: Western Blot
    Sample Tested: mouse brain, Brain olfactory membrane fraction and Adult brain (hippocampus)
    Species: Mouse
    Verified Customer | Posted 04/15/2021
    This antibody is working great without non-specific bands.
    Human/Mouse NCAM‑1/CD56 Antibody AF2408
  • Human/Mouse NCAM-1/CD56 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Mouse brain cancer sample and Mouse brain cancer tissue
    Species: Mouse
    Verified Customer | Posted 01/19/2017
    Immunohistochemistry-Floating section

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