Human/Mouse NCAM-1/CD56 Antibody

Catalog # Availability Size / Price Qty
AF2408
AF2408-SP
Detection of Human and Mouse NCAM‑1/CD56 by Western Blot.
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Product Details
Citations (9)
FAQs
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Reviews (2)

Human/Mouse NCAM-1/CD56 Antibody Summary

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 either lyophilized or 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
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.
 

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

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

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 antibody in SH-SY5Y Human Neuroblastoma Cells by Immunocytochemistry (ICC). View Larger

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.

Immunocytochemistry NCAM-1/CD56 antibody in BG01V Human Embryonic Stem Cells by Immunocytochemistry (ICC). View Larger

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.

Simple Western Detection of Human and Mouse NCAM-1/CD56 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Knockout Validated View Larger

Western Blot Shows Human NCAM‑1/CD56 Specificity by Using Knockout Cell Line. Western blot shows lysates of U937 human histiocytic lymphoma cell line and human NCAM-1 knockout U937 human histiocytic lymphoma cell line (KO). PVDF membrane was probed with 0.25 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 (HAF017). A specific band was detected for NCAM‑1/CD56 at approximately 160 kDa (as indicated) in the parental U937 human histiocytic lymphoma cell line, but is not detectable in knockout U937 human histiocytic lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human/Mouse NCAM-1/CD56 Antibody

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.

9 Citations: Showing 1 - 9
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  1. Abnormal ciliogenesis in decidual stromal cells in recurrent miscarriage
    Authors: E Hassan, R Kojima, F Ozawa, H Yoshihara, S Goto, T Kitaori, H Inagaki, Y Kato, M Sugiura-Og
    Journal of reproductive immunology, 2022;150(0):103486.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Description of Osmolyte Pathways in Maturing Mdx Mice Reveals Altered Levels of Taurine and Sodium/Myo-Inositol Co-Transporters
    Authors: C Merckx, G Cosemans, J Zschüntzsc, R Raedt, J Schmidt, B De Paepe, JL De Bleecke
    International Journal of Molecular Sciences, 2022;23(6):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  3. CD200S-positive granulated lymphoid cells in endometrium appear to be CD56-positive uterine NK cells
    Authors: DA Clark, A Reihani, JL Arredondo, K Ask, WG Foster
    Journal of reproductive immunology, 2022;150(0):103477.
    Species: Human
    Sample Types: Whole Tissues
    Applications: IHC
  4. Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Authors: W Kim, H Watanabe, S Lomoio, G Tesco
    The Journal of Biological Chemistry, 2021;0(0):100372.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  5. Mapping Gene Circuits Essential for Germ Layer Differentiation via Loss-of-Function Screens in Haploid Human Embryonic Stem Cells
    Authors: A Yilmaz, C Braverman-, A Bialer-Tsy, M Peretz, N Benvenisty
    Cell Stem Cell, 2020;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  6. Generation of three iPSC lines from two patients with heterozygous FOXF1 mutations associated to Alveolar Capillary Dysplasia with Misalignment of the Pulmonary Veins
    Authors: E Slot, A de Klein, RJ Rottier
    Stem Cell Res, 2020;44(0):101745.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  7. Tumor suppressor TET2 promotes cancer immunity and immunotherapy efficacy
    Authors: YP Xu, L Lv, Y Liu, MD Smith, WC Li, XM Tan, M Cheng, Z Li, M Bovino, J Aubé, Y Xiong
    J. Clin. Invest., 2019;130(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  8. 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 Types: Whole Cells
    Applications: ICC
  9. 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 Types: Cell Lysates
    Applications: Western Blot

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Reviews for Human/Mouse NCAM-1/CD56 Antibody

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Human/Mouse NCAM-1/CD56 Antibody
By WonHee Kim on 04/15/2021
Application: Western Blot Sample Tested: Adult brain (hippocampus),Brain olfactory membrane fraction,mouse brain Species: Mouse

This antibody is working great without non-specific bands.


Human/Mouse NCAM-1/CD56 Antibody
By Anonymous on 01/19/2017
Application: Immunohistochemistry Sample Tested: Mouse brain cancer sample,Mouse brain cancer tissue Species: Mouse

Immunohistochemistry-Floating section