CNTFR alpha Antibody [Unconjugated]

Recombinant Rat CNTF R alpha (Catalog # 558-CR) enhances Recombinant Rat CNTF (Catalog # 557-NT) induced proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Enhancement of Recombinant Rat CNTF (10 ng/mL) activity elicited by Recombinant Rat CNTF R alpha (0.25 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat CNTF R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑559‑NA). The ND50 is typically 2‑6 µg/mL.

CNTF activates tyrosine hydroxylase by phosphorylation in mouse and human locus coeruleusCntfr mRNA expression in mouse locus coeruleus (LC, white arrows). Increasing mRNA content is indicated by a color gradient ranging from dark blue (low) to red (high). The ependymal surface as contact point with the 4th ventricle is shown in blue. Scale bar = 100 μm.Hypothetical orientation of NE neurons with their dendrites emanating toward the 4th ventricle. (B1) CNTF receptor (CNTFR) localization in tyrosine hydroxylase (TH)+ processes extending toward the ventricular wall (arrow marks ventricular surface). Scale bar = 10 μm.c‐Fos activation in LC 2 h after formalin stress. Left: Increased density of c‐Fos+ cells in the LC. Scale bar = 150 μm. Right: Quantitative data. *P 4 mice/genotype.Data information: Data are expressed as means ± s.e.m. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30209240), licensed under a CC-BY license. Not internally tested by R&D Systems.

Secretagogin expression in rodent brain stem (related to Fig 4)A, A1Labeling specificity of our anti‐secretagogin antibody was verified in Scgn−/− mice (A1). These experiments also showed that tyrosine hydroxylase (TH)+ LC neurons (*) co‐express secretagogin in wild‐type (solid arrowheads) but not null mice (open arrowheads). Scale bars = 200 μm (low‐power micrographs, left) and 10 μm (high‐resolution insets, right).BSecretagogin+ neurons populate the locus coeruleus (LC) in the rat. Solid red circles pinpoint the general location of perikarya, while fine lines correspond to axons coursing toward the surface (4th ventricle).C, C1Secretagogin was expressed in both the somata (arrowheads; C) and dendrites (arrowheads; C1) of most TH+ neurons in the rat, too. Scale bars = 30 μm.DSecretagogin is localized to axons (arrowheads) in the rat. (D) Dorsal nucleus of vagus (nX), “2” shows the nucleus at high resolution. Scale bars = 200 μm (D), 30 μm (2).ESecretagogin (SCGN) is enriched in the synaptosomal fraction prepared from the mouse prefrontal cortex, lending additional support for its axonal localization. Synaptophysin (SYP) was used to control the enrichment of presynaptic proteins.FSecretagogin+ neurons can co‐express neuropeptide Y (NPY) in the LC of the rat. Open and solid arrowheads indicate the lack and existence of NPY co‐localization in adjacent neurons, respectively. Scale bars = 10 μm.GRepresentative Western blots from Cntf−/−, Scgn−/−, and wild‐type mice with or without intracerebroventricular CNTF treatment.H, H1INS‐1E cells were used as a cellular model exclusively because of their native co‐expression of CNTF receptor‐ alpha (CNTFR alpha ), tyrosine hydroxylase (TH), secretagogin (SCGN), and extracellular signal‐regulated kinase 1/2 (Erk1/2). Red arrows label target proteins at their predicted sizes. Multiple bands likely represent splice variants, posttranslational modifications, or multimers. Antibodies were also validated histochemically often using knock‐out tissues.ISecretagogin, phospho‐Ser31‐TH, phospho‐Erk1, and total Erk1 expression in INS‐1E cells stimulated with CNTF with/without concomitant silencing of secretagogin (Scgn) gene expression (for 48 h). Note the lack of Erk1 phosphorylation upon genetic inactivation of secretagogin.JSignificant reduction of secretagogin (SCGN) protein levels upon acute RNAi‐mediated gene silencing in vitro (for 48 h). CNTF exposure did not affect the efficacy of gene silencing. Experiments were performed in triplicate; *P 4 mice/genotype.Data information: Data are expressed as means ± s.e.m. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30209240), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Rt
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CNTFR alpha Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant rat CNTF R alpha
  Ala19-Pro346
  Accession # Q08406
• Specificity: Detects human CNTF R alpha in direct ELISAs and Western blots. In direct ELISAs, greater than 50% cross-reactivity with recombinant human CNTF sR alpha is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Cntfr
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 2-6 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. *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: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for CNTFR alpha Antibody [Unconjugated]

• Ciliary neurotrophic factor receptor subunit alpha;Cntfr
• CNTF R alpha
• CNTFR alpha
• CNTFR
• CNTFRa

Background

The high-affinity CNTF receptor complex, which mediates the biological action of CNTF, contains three proteins: the ligand-binding a subunit (CNTF R alpha ) and the two signal-transducing proteins LIF R beta and gp130. Whereas LIF R beta and gp130 are widely expressed in many cell types, the expression of CNTF R alpha is restricted to the central and peripheral nervous systems. cDNAs encoding CNTF R alpha have been isolated from both human and rat and were shown to share 94% amino acid (aa) sequence identity. Rat CNTF R alpha cDNA encodes a 372 amino acid residue precursor protein that apparently has a 22 aa residue signal peptide and five potential glycoslyation sites. CNTF R alpha differs from other cytokine receptors in that it lacks transmembrane and cytoplasmic domains and is anchored to cell membranes by a glycosylphosphatidylinositol (GPI) linkage. Similar to other GPI-linked proteins, soluble CNTF receptor alpha (CNTF sR alpha ) can be released from the cell surface by phosphatidylinositol-specific phospholipase C. CNTF sR alpha can be released from skeletal muscle in response to peripheral nerve injury and high concentrations of CNTF sR alpha have also been detected in human cerebrospinal fluid. CNTF sR alpha binds CNTF in solution and the complex can act on cells that express only LIF R beta and gp130 but not CNTF R alpha .

1. Yancopoulos, G.D. in Guidebook to Cytokines and Their Receptors,
Nicola, N.A. editor, Oxford University Press, New York, pp137.

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 CNTFR alpha Antibody (AF-559-NA)(3)

Publications using AF-559-NA

• P Gómez-Fern, A Lopez de L, I Astobiza, J Mena, A Urtasun, V Altmann, F Matesanz, D Otaegui, E Urcelay, A Antigüedad, S Malhotra, X Montalban, T Castillo-T, L Espino-Pai, O Aktas, M Buttmann, A Chan, B Fontaine, PA Gourraud, M Hecker, S Hoffjan, C Kubisch, T Kümpfel, F Luessi, UK Zettl, F Zipp, I Alloza, M Comabella, CM Lill, K Vandenbroe The Rare IL22RA2 Signal Peptide Coding Variant rs28385692 Decreases Secretion of IL-22BP Isoform-1, -2 and -3 and Is Associated with Risk for Multiple Sclerosis Cells, 2020-01-10;9(1):. 2020-01-10 [PMID: 31936765] (Western Blot, Mouse)
• Alán Alpár, Péter Zahola, János Hanics, Zsófia Hevesi, Solomiia Korchynska, Marco Benevento et al. Hypothalamic CNTF volume transmission shapes cortical noradrenergic excitability upon acute stress The EMBO Journal 2018-11-02 [PMID: 30209240]
• Nancy Lee, Carolyn E. Rydyznski, Rachel P. Spearry, Rachel Robitz, A. John MacLennan The contribution of ciliary neurotrophic factor receptors to adult motor neuron survival in vivo is specific to insult type and distinct from that for embryonic motor neurons Journal of Comparative Neurology 2013-10-01 [PMID: 23695797] (Immunohistochemistry, Transgenic Mouse)

Bioinformatics

• Gene Symbol: Cntfr
• Uniprot:
  • Q08406()