NCAM-1/CD56 Antibody (809220) [Unconjugated]

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.

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.

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.

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.

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.

• Reactivity: Mu
• Applications: WB, Flow, IHC, CyTOF-ready, mIF
• Clone: 809220
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: LYOPH

NCAM-1/CD56 Antibody (809220) [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: NCAM1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 3-25 ug/mL
  • Multiplex Immunofluorescence 15 ug/mL
  • Western Blot 0.1 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. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for NCAM-1/CD56 Antibody (809220) [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

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.

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 (MAB7820)(2)

Publications using MAB7820

• X Wang, M Lu, X Tian, Y Ren, Y Li, M Xiang, S Chen Diminished expression of major histocompatibility complex facilitates the use of human induced pluripotent stem cells in monkey Stem Cell Res Ther, 2020-08-03;11(1):334. 2020-08-03 [PMID: 32746912] (Western Blot, Human)
• EM Treutlein, K Kern, A Weigert, N Tarighi, CD Schuh, RM Nüsing, Y Schreiber, N Ferreirós, B Brüne, G Geisslinge, S Pierre, K Scholich The prostaglandin E2 receptor EP3 controls CC-chemokine ligand-mediated neuropathic pain induced by mechanical nerve damage J. Biol. Chem., 2018-05-11;0(0):. 2018-05-11 [PMID: 29752406] (Flow Cytometry, Mouse)

Bioinformatics

• Gene Symbol: NCAM1
• Uniprot:
  • P13595()