14-9118-80

antibody from Invitrogen Antibodies

Targeting: NCAM1 CD56, NCAM

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-9118-80

Provider

Invitrogen Antibodies

Product name

PSA-NCAM Monoclonal Antibody (12E3), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The monoclonal antibody 12E3 specifically recognizes polysialylated neural cell adhesion molecule (PSA-NCAM) in human, mouse and rat. Polysialic acid is a long homopolymer of sialic acid that is negatively charged and is attached to neural cell adhesion molecule (CD56) and serves as a regulator of NCAM function. PSA attachment to NCAM is associated with inhibited adhesion of neural cells. PSA-NCAM expression is highly regulated and corresponds to specific neural developmental windows in which neural precursors are migrating and during the process of axonal sprouting, guidance, and targeting. PSA-NCAM expression is prevalent during development of the brain, but in the adult becomes restricted to regions undergoing self-renewal or exhibiting plasticity such as the olfactory bulb, suprachiasmatic nucleus, hippocampus, hypothalamus, and specific spinal cord nuclei. PSA-NCAM is re-expressed during tumorigenesis and is also expressed on cell lines isolated from neuroblastomas and pheochromocytomas. Applications Reported: This 12E3 antibody has been reported for use in immunohistochemical staining of frozen tissue sections, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, microscopy, and immunocytochemistry. Applications Tested: This 12E3 antibody has been tested by immunocytochemistry of formaldehyde-fixed PC-12 (rat) cells and can be used at less than or equal to 20 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgM

Antibody clone number

12E3

Vial size

25 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Neurogenesis of medium spiny neurons in the nucleus accumbens continues into adulthood and is enhanced by pathological pain.

García-González D, Dumitru I, Zuccotti A, Yen TY, Herranz-Pérez V, Tan LL, Neitz A, García-Verdugo JM, Kuner R, Alfonso J, Monyer H
Molecular psychiatry 2021 Sep;26(9):4616-4632

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Arginase Inhibition Supports Survival and Differentiation of Neuronal Precursors in Adult Alzheimer's Disease Mice.

Polis B, Srikanth KD, Gurevich V, Bloch N, Gil-Henn H, Samson AO
International journal of molecular sciences 2020 Feb 8;21(3)

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Bisecting GlcNAc Is a General Suppressor of Terminal Modification of N-glycan.

Nakano M, Mishra SK, Tokoro Y, Sato K, Nakajima K, Yamaguchi Y, Taniguchi N, Kizuka Y
Molecular & cellular proteomics : MCP 2019 Oct;18(10):2044-2057

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Oxidative Stress-Tolerant Stem Cells from Human Exfoliated Deciduous Teeth Decrease Hydrogen Peroxide-Induced Damage in Organotypic Brain Slice Cultures from Adult Mice.

Xiao L, Saiki C, Okamura H
International journal of molecular sciences 2019 Apr 15;20(8)

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Human Dental Pulp Cells Differentiate toward Neuronal Cells and Promote Neuroregeneration in Adult Organotypic Hippocampal Slices In Vitro.

Xiao L, Ide R, Saiki C, Kumazawa Y, Okamura H
International journal of molecular sciences 2017 Aug 11;18(8)

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Two separate subtypes of early non-subplate projection neurons in the developing cerebral cortex of rodents.

Espinosa A, Gil-Sanz C, Yanagawa Y, Fairén A
Frontiers in neuroanatomy 2009;3:27

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Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer.

Wakayama T, Tabar V, Rodriguez I, Perry AC, Studer L, Mombaerts P
Science (New York, N.Y.) 2001 Apr 27;292(5517):740-3

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Removal of polysialic acid-neural cell adhesion molecule induces aberrant mossy fiber innervation and ectopic synaptogenesis in the hippocampus.

Seki T, Rutishauser U
The Journal of neuroscience : the official journal of the Society for Neuroscience 1998 May 15;18(10):3757-66

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Highly polysialylated neural cell adhesion molecule (NCAM-H) is expressed by newly generated granule cells in the dentate gyrus of the adult rat.

Seki T, Arai Y
The Journal of neuroscience : the official journal of the Society for Neuroscience 1993 Jun;13(6):2351-8

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-PSA-NCAM Monoclonal Antibody (12E3), eBioscience(Product # 14-9118-82) and a ~150kDa band corresponding to PSA-NCAM was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), IMR-32 (Lane 2), NK-92 (Lane 3), MCF7 (Lane 4), Hep G2 (Lane 5), Neuro-2a (Lane 6), PC-12 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry of fixed PC-12 cells stained with 20 µg/mL of Mouse IgM Isotype Control Purified (left) or 20 µg/mL of Anti-Polysialylated Neural Cell Adhesion Molecule (PSA-NCAM) Purified (right), followed by Anti-mouse IgM Biotin and Streptavidin FITC.Nuclei are stained with DAPI.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of PSA-NCAM was performed using 70% confluent log phase IMR-32 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with PSA-NCAM Monoclonal Antibody (12E3), eBioscience (Product # 14-9118-82) at 20 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Plasma membrane localization. Panel e represents HepG2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Effect of DPCs on neurogenesis in adult mouse hippocampal slices in vitro. ( A ) Experimental setup. Mouse hippocampus slices were cultivated on matrigel with DPCs or by themselves for 11 days, followed by immunofluorescence staining; ( B ) Images were taken by a phase contrast microscope. Hippocampus slice-derived cells showed glia cell-like morphology when cultivated alone (left). However, when the hippocampus slice co-cultivated with DPCs, the derived cells exhibited neuron-like morphology and formed networks. Scale bar = 50 um ( C ) Immunofluorescence staining showed that cells derived from the co-cultivated hippocampus slice are positive to the mature neuron mark NeuN (right). However, cells derived from the single cultivated hippocampus did not react with NeuN (left). Images were taken by LSM. Scale bar = 25 um; ( D ) In solo or co-cultivated hippocampus slices, cells reacted with NeuN and PSA-NCAM (a marker of developing and migrating neurons) antibodies. Images were taken by LSM. Scale bar = 50 um.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 2 Representative x40 bright-field micrographs of the hippocampal dentate gyri of 3 x Tg mice with x100 insets ( A , B ). The subgranular zone (SGZ) located polysialylated neuronal cell adhesion molecule (PSA-NCAM) positive cells are marginally present in vehicle-treated animals ( A ) but show much greater incidence in norvaline-treated mice with penetration into the granule cell layer ( B ). The treatment is associated with a significant increase in the PSA-NCAM immunopositive area ( C ) and stain intensity ( D ). Scale bars 50 um, insets 10 um. The data are presented as means +- SEM. * p < 0.05, ** p < 0.01, (two-tailed Student's t -test), n = 8.