Human NCAM‑1/CD56 Antibody

Name: Human NCAM‑1/CD56 Antibody
Brand: R&D Systems
SKU: MAB24081
Price: 489 USD
Availability: InStock

R&D Systems | Catalog # MAB24081

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Citations (15)

Product Documents

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rabbit

Applications

Validated:

Multiplex Immunofluorescence, Immunohistochemistry, Western Blot, Flow Cytometry, COMET, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2B Clone # 301021

View all NCAM-1/CD56 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human NCAM-1/CD56
Leu20-Pro603 and Glu636-Asn741
Accession # NP_001070150

Specificity

Detects human NCAM-1/CD56 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) ALCAM, rhBCAM, rhEPCAM, rhMCAM, rhNCAM-1-L1, recombinant mouse (rm) MAdCAM-1, rmNCAM-1-L1, or rmOCAM is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

Scientific Data Images for Human NCAM‑1/CD56 Antibody

Detection of NCAM-1/CD56 in Human Brain Cortex via seqIF™ staining on COMET™

NCAM-1/CD56 was detected in immersion fixed paraffin-embedded sections of human brain Cortex using Mouse Anti-Human NCAM-1/CD56, Monoclonal Antibody (Catalog # MAB24081) at 25ug/mL at 37° Celsius for 8 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the neuropil. Protocol available in COMET™ Panel Builder.

Detection of Human NCAM‑1/CD56 by Western Blot.

Western blot shows lysates of human brain (cerebellum and hypothalamus). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human NCAM‑1/CD56 Monoclonal Antibody (Catalog # MAB24081) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for NCAM‑1/CD56 at approximately 105-130 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of NCAM-1/CD56 in Human PBMC by Flow Cytometry.

Human PBMCs were stained with either (A) Mouse Anti-Human NCAM-1/CD56 Monoclonal Antibody (Catalog # MAB24081) or (B) Mouse IgG2B Isotype Control (MAB0041) followed by Anti-Mouse IgG APC-conjugated secondary antibody (F0101B) and Mouse Anti-Human NKp46 PE-conjugated Monoclonal Antibody (FAB1850P). Staining was performed using our Staining Membrane-associated Proteins protocol.

Detection of NCAM-1/CD56 in Human NK cells by Flow Cytometry.

Human NK cells were expanded from PBMCs using Cloudz Human NK Cell Expansion Kit (CLD004) and were stained with either (A) Mouse Anti-Human NCAM-1/CD56 Monoclonal Antibody (Catalog # MAB24081) or (B) Mouse IgG2B Isotype Control (MAB0041) followed by Anti-Mouse IgG APC-conjugated secondary antibody (F0101B) and Mouse Anti-HumanCD3 PE-conjugated Monoclonal Antibody (FAB100P). Staining was performed using our Staining Membrane-associated Proteins protocol.

Detection of NCAM‑1/CD56 in Human Brain Cortex.

NCAM‑1/CD56 was detected in immersion fixed paraffin-embedded sections of human brain cortex using Mouse Anti-Human NCAM‑1/CD56 Monoclonal Antibody (Catalog # MAB24081) at 25 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001) or the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the neuropil. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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Applications for Human NCAM‑1/CD56 Antibody

Application

Recommended Usage

COMET

Optimal dilutions of this antibody should be experimentally determined.

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 and human NK cells expanded from PBMCs using Cloudz Human NK Cell Expansion Kit (Catalog # CLD004)

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cortex)

Multiplex Immunofluorescence

25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human Brain Cortex

Western Blot

2 µg/mL
Sample: Human brain (cerebellum and hypothalamus)

Flow Cytometry Panel Builder

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Advanced Features

Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
Spillover Popups - Visualize the spectra of individual fluorochromes
Antigen Density Selector - Match fluorochrome brightness with antigen density

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Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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).