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Human NCAM-1/CD56 MAb (Clone 301021) (25 ug)
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咨询Flow Cytometry(0.25 µg/106 cells)
Immunohistochemistry(8-25 µg/mL)
Leu20-Pro603 and Glu636-Asn741
Accession # NP_001070150
Human NCAM-1/CD56 Antibody Summary
Leu20-Pro603 and Glu636-Asn741
Accession # NP_001070150
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Applications
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.
Scientific Data
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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.
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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.
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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.
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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.
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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.
Reconstitution Calculator
Preparation and Storage
- 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).
- Burg, M.A. et al. (1995) J. Neurosci. Res. 41:49.
- Storms, S.D. and U. Rutishauser (1998) J Biol. Chem. 273:27124.
- Margolis, R.K. et al. (1996) Perspect. Dev. Neurobiol. 3:273.
- Dickson, G. et al. (1987) Cell 50:1119.
- Lanier, L.L. et al. (1991) J. Immunol. 146:4421.
- Hemperly, J.J. et al. (1990) J. Mol. Neurosci. 2:71.
- Rutishauser, U.and C. Goridis (1986) Trends Genet. 2:72.
- Vawter, M.P. et al. (2001) Exp. Neurol. 172:29.
- Rutihauser, U. (1990) Adv. Exp. Med. Biol. 265:179.
- Becker, C.G. et al. (1996) J. Neurosci. Res. 45:143.
- Doherty, P. et al. (1995) J. Neurobiol. 26:437.
- Eckardt, M. et al. (2000) J. Neurosci. 20:5234.
- Muller, D. et al. (1996) Neuron 17:413.
Product Datasheets
Flow Cytometry(0.25 µg/106 cells)
Immunohistochemistry(8-25 µg/mL)
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).
- Burg, M.A. et al. (1995) J. Neurosci. Res. 41:49.
- Storms, S.D. and U. Rutishauser (1998) J Biol. Chem. 273:27124.
- Margolis, R.K. et al. (1996) Perspect. Dev. Neurobiol. 3:273.
- Dickson, G. et al. (1987) Cell 50:1119.
- Lanier, L.L. et al. (1991) J. Immunol. 146:4421.
- Hemperly, J.J. et al. (1990) J. Mol. Neurosci. 2:71.
- Rutishauser, U.and C. Goridis (1986) Trends Genet. 2:72.
- Vawter, M.P. et al. (2001) Exp. Neurol. 172:29.
- Rutihauser, U. (1990) Adv. Exp. Med. Biol. 265:179.
- Becker, C.G. et al. (1996) J. Neurosci. Res. 45:143.
- Doherty, P. et al. (1995) J. Neurobiol. 26:437.
- Eckardt, M. et al. (2000) J. Neurosci. 20:5234.
- Muller, D. et al. (1996) Neuron 17:413.
Preparation and Storage
- 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.