Perfusion fixed frozen sections of rat brain (cortex)
Measured by its ability to neutralize CNTF R alpha -enhanced proliferation in the TF‑1 human erythroleukemic cell line. The Neutralization Dose (ND50) is typically 2-6 µg/mL in the presence of 0.25 µg/mL Recombinant Rat CNTF R alpha and 10 ng/mL Recombinant Rat CNTF.
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.
CNTF R alpha Enhancement of CNTF-dependent Cell Proliferation and Neutralization by Rat CNTF R alpha Antibody. 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
Background: CNTF R alpha
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.
Yancopoulos, G.D. in Guidebook to Cytokines and Their Receptors,
Nicola, N.A. editor, Oxford University Press, New York, pp137.
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