Contactin-2 (CNTN2), also called TAG1 (transient axonal glycoprotein), TAX1 (transiently-expressed axonal glycoprotein), or axonin-1, is a 135 kDa glycosyl-phosphatidylinositol (GPI)- anchored cell adhesion molecule that belongs to the contactin subfamily within the immunoglobulin (Ig) protein superfamily (1-3). Mouse Contactin-2 cDNA encodes a 30 amino acid (aa) signal peptide, a 984 aa mature secreted protein with 6 Ig-like domains followed by 4 fibronectin type III-like repeats, and a 26 aa C-terminal GPI anchor pro-sequence. GPI-specific phospholipase activity can release soluble, active Contactin-2 from the membrane (2). Mature mouse Contactin-2 shares approximately 93%, 97%, and 77% aa sequence identity with human, rat, and chicken Contactin-2, respectively. During development, Contactin-2 is expressed by a subset of neuronal populations in the central nervous system (CNS) and peripheral nervous system (PNS), particularly during initial phases of axon outgrowth (3-5). Both the 135 kDa form and a 90 kDa form are also upregulated in response to CNS injury in the adult (6). Data support a role for Contactin-2 in axon pathfinding, neurite outgrowth and adhesion, especially in the CNS (3-6). In mature myelinated fibers, Contactin-2 is expressed by oligodendrocytes and Schwann cells, which are myelinating glial cells of the CNS and PNS, respectively (7, 8). It is enriched in the juxtaparanodal regions, where it recruits contactin-associated protein 2 (caspr2), a transmembrane neurexin involved in cell adhesion and intercellular communication (7-10). The axonal Contactin-2 interacts in cis with caspr2 and in trans with another Contactin-2 on the glial membrane (8). This ternary complex is required for the accumulation and organization of K+ channels in the juxtaparanodes (9).
Human/Mouse/Rat Contactin‑2/TAG1 Antibody
R&D Systems | Catalog # AF4439
Key Product Details
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Human, Mouse, Rat, Hamster - Mesocricetus auratus (Golden Hamster), Primate, Transgenic Mouse
Applications
Validated:
Immunohistochemistry, Western Blot, Simple Western
Cited:
Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, Mass Spectrometry
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Contactin-2/TAG1
Gln31-Ser1014
Accession # Q61330
Gln31-Ser1014
Accession # Q61330
Specificity
Detects mouse and rat Contactin-2 in direct ELISAs. Detects human, mouse, and rat Contactin-2 in Western blots.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human/Mouse/Rat Contactin‑2/TAG1 Antibody
Detection of Human, Mouse, and Rat Contactin‑2/TAG1 by Western Blot.
Western blot shows lysates of human cerebellum tissue, mouse brain tissue, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat Contactin-2/TAG1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4439) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Contactin-2/TAG1 at approximately 135 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Contactin‑2/TAG1 in Mouse Embryo.
Contactin-2/TAG1 was detected in immersion fixed frozen sections of mouse embryo (E13) using Goat Anti-Human/Mouse/Rat Contactin-2/TAG1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4439) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to muscle cells in proximity to ribs. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.Detection of Mouse Contactin‑2/TAG1 by Simple WesternTM.
Simple Western lane view shows lysates of mouse spinal cord tissue, loaded at 0.2 mg/mL. A specific band was detected for Contactin-2/TAG1 at approximately 162 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat Contactin-2/TAG1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4439) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.Detection of Mouse Human/Mouse/Rat Contactin-2/TAG1 Antibody by Western Blot
Quantitative proteomics in Rab35 cKO P0 hippocampus.a Volcano plot of the TMT-based quantitative proteomes identifying the dysregulated proteins in Rab35 cKO hippocampus in comparison with the control hippocampus (n = 5 mice per genotype). b Number of proteins identified as significantly dysregulated and as either membrane traffic-related or neuronal migration-related. c, d Western blot analysis of control and Rab35 cKO P0 hippocampi using anti-contactin-2, anti-CHL1, and anti-actin antibodies. e, f Quantification of contactin-2 (c) and CHL1 (d) protein levels in control and Rab35 cKO P0 hippocampi. Band intensities of the indicated proteins were normalized to those of actin (n = 9 mice per genotype). Unpaired Student’s t-test; e, p = 0.0153; f, p = 0.0095. g, h Levels of contactin-2 (g) and CHL1 (h) were quantified by targeted MS using the PRM method (n = 5 mice per genotype). Unpaired Student’s t-test; gp = 0.0053; hp = 0.0229. i Western blot analysis of control and Rab35 cKO P0 hippocampus using anti-N-cadherin and anti-actin antibodies. j Quantification of N-cadherin protein levels in the control and Rab35 cKO P0 hippocampus (n = 9 mice per genotypes). Unpaired Student’s t-test, p = 0.9020. k Representative images of DIV 2 hippocampal primary neurons stained for contactin-2 (green), rhodamine-phalloidin (magenta) and DAPI (blue). Scale bar, 20 μm. l Quantification of contactin-2 intensity at the somatic plasma membrane in control (n = 4) and Rab35-deficient (n = 4) cells. Thirty neurons from four different cultures per genotype were analyzed. Mann–Whitney U-test, p = 0.0286. Data represent the mean ± SEM; n.s. not significant (p > 0.05); *p < 0.05; **p < 0.01. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/37085665), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human/Mouse/Rat Contactin‑2/TAG1 Antibody
Application
Recommended Usage
Immunohistochemistry
5-15 µg/mL
Sample:
Sample:
Immersion fixed frozen sections of mouse embryo (E13)
Simple Western
10 µg/mL
Sample: Mouse spinal cord tissue
Sample: Mouse spinal cord tissue
Western Blot
1 µg/mL
Sample: Human cerebellum tissue, Mouse brain tissue, and Rat brain tissue
Sample: Human cerebellum tissue, Mouse brain tissue, and Rat brain tissue
Reviewed Applications
Read 2 reviews rated 4.5 using AF4439 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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
Background: Contactin-2/TAG1
References
- Wolfer, D. and R.J. Giger (1994) Swissprot Accession # Q61330.
- Hasler, T.H. et al. (1993) Eur. J. Biochem. 211:329.
- Karagogeos, D. (2003) Front. Biosci. 8:s1304.
- Liu, Y. and M.C. Halloran (2005) J. Neurosci. 25:10556.
- Denaxa, M. et al. (2005) Dev. Biol. 288:87.
- Soares, S. et al. (2005) Eur. J. Neurosci. 21:1169.
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Traka, M. et al. (2002) J. Neurosci. 22:3016.
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Poliak, S. and E. Peles (2003) Nat. Rev. Neurosci. 4:968.
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Traka, M. et al. (2003) J. Cell Biol. 162:1161.
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Poliak, S. et al. (2003) J. Cell Biol. 162:1149.
Alternate Names
Axonin-1, AXT, CNTN2, Contactin2, TAG1, TAX1
Gene Symbol
CNTN2
UniProt
Additional Contactin-2/TAG1 Products
Product Documents for Human/Mouse/Rat Contactin‑2/TAG1 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse/Rat Contactin‑2/TAG1 Antibody
For research use only
Related Research Areas
Citations for Human/Mouse/Rat Contactin‑2/TAG1 Antibody
Customer Reviews for Human/Mouse/Rat Contactin‑2/TAG1 Antibody (2)
4.5 out of 5
2 Customer Ratings
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Application: ImmunohistochemistrySample Tested: mouse heartSpecies: MouseVerified Customer | Posted 08/05/2020
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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