Mouse NCAM-1/CD56 Antibody (809220)

Name: Mouse NCAM-1/CD56 Antibody (809220)
Brand: R&D Systems
SKU: MAB7820
Price: 449 USD
Availability: InStock

R&D Systems | Catalog # MAB7820

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Citations (4)

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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:

Mouse

Cited:

Human, Mouse

Applications

Validated:

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

Cited:

Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG_2A Clone # 809220

View all NCAM-1/CD56 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse NCAM‑1/CD56
Leu20-Thr711
Accession # P13595

Specificity

Detects mouse NCAM‑1/CD56 in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Rat

Isotype

IgG_2A

Scientific Data Images for Mouse NCAM-1/CD56 Antibody (809220)

Detection of CD56/NCAM1 in Mouse Cerebellum via seqIF™ staining on COMET™

CD56/NCAM1 was detected in immersion fixed paraffin-embedded sections of mouse Cerebellum using Rat Anti-Mouse CD56, Monoclonal Antibody (Catalog # MAB7820) at 15ug/mL at 37° Celsius for 4 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-Rat IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the neuropil. Protocol available in COMET™ Panel Builder.

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

Western blot shows lysates of Neuro-2A mouse neuroblastoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Mouse NCAM-1/CD56 Monoclonal Antibody (Catalog # MAB7820) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for NCAM-1/CD56 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of NCAM‑1/CD56 in Neuro‑2A Mouse Cell Line by Flow Cytometry.

Neuro-2A mouse neuroblastoma cell line was stained with Rat Anti-Mouse NCAM-1/CD56 Monoclonal Antibody (Catalog # MAB7820, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). View our protocol for Staining Membrane-associated Proteins.

Detection of NCAM‑1/CD56 in Mouse Cerebellum.

NCAM‑1/CD56 was detected in immersion fixed paraffin-embedded sections of mouse cerebellum using Rat Anti-Mouse NCAM‑1/CD56 Monoclonal Antibody (Catalog # MAB7820) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC005). 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 cytoplasm and neuropils. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of NCAM‑1/CD56 in Mouse Cortex.

NCAM‑1/CD56 was detected in immersion fixed paraffin-embedded sections of mouse cortex using Rat Anti-Mouse NCAM‑1/CD56 Monoclonal Antibody (Catalog # MAB7820) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC005). 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 cytoplasm and neuropils. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

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Applications for Mouse NCAM-1/CD56 Antibody (809220)

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: Neuro‑2A mouse neuroblastoma cell line

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of mouse cerebellum and mouse cortex

Multiplex Immunofluorescence

15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of mouse cerebellum

Western Blot

0.1 µg/mL
Sample: Neuro‑2A mouse neuroblastoma cell line

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

Sterile PBS to a final concentration of 0.5 mg/mL. 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.<br/>*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

NCAM-1 (Neural adhesion molecule-1; also CD56) is a 120-190 kDa glycoprotein member of the Ig Superfamily. It is expressed on multiple cell types, both in the embryo and adult. Here, it serves as both an adhesion molecule and a receptor for multiple ligands, including as FGFR, PDGF, GDNF and agrin. On the cell surface, it is a cis-oriented homodimer that can form homodimers in-trans with other cis-homodimers. In the embryo, NCAM-1 is polysialylated (PolySia), and shows a MW of 200-220 kDa in SDS-PAGE. This polysialylation reduces the ability of NCAM-1 to dimerize. Mature mouse NCAM-1 is a 1096 amino acid (aa) type I transmembrane (TM) protein (aa 20-1115). It possesses a 692 aa extracellular region (aa 20-711) and a 386 aa cytoplasmic domain. The extracellular region contains five consecutive C2-type Ig-like domains (aa 20-492) followed by two FN type-III domains (aa 497-692). Multiple splice variants exist. There is a 140 kDa TM variant that shows a deletion of aa 810-1076, and a 120 kDa variant that is GPI-linked and shows a 24 aa substitution for aa 702-1115. A third potential variant contains a five aa substitution for aa 601-1115. Over aa 20-711, mouse NCAM-1 shares 99% and 95% aa identity with rat and human NCAM-1, respectively.