Human Nogo Receptor/RTN4R HEK293 Overexpression Lysate

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Human Nogo Receptor/RTN4R HEK293 Overexpression Lysate: Product Information

Product Description
This Human Nogo Receptor/RTN4R overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Nogo Receptor/RTN4R protein (Cat: 10466-H03H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the mature form of human RTN4R (NP_075380.1) (Met 1-Ser 447) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human RTN4R/Fc is a disulfide-linked homodimer. The reduced monomer consists of 669 amino acids and has a predicted molecular mass of 73 kDa. As a result of glycosylation, the apparent molecular mass of rh RTN4R/Fc monomer is approximately 100 kDa in SDS-PAGE under reducing conditions.

Human Nogo Receptor/RTN4R HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human Nogo Receptor/RTN4R HEK293 Overexpression Lysate: Alternative Names

Human NGR Overexpression Lysate; Human NOGOR Overexpression Lysate

Nogo Receptor/RTN4R Background Information

Reticulon 4 receptor (RTN4R), also known as Nogo-66 Receptor (NgR), is a glycosylphosphoinositol (GPI)-anchored protein that belongs to the Nogo recptor family including three members. Mouse RTN4R cDNA contains 1 LRP (Leucine-rich) repeats. RTN4R is expressed predominantly in neurons and their axons in the central nervous systems (CNS). As a receptor for myelin-derived proteins Nogo, myelin-associated glycoprotein (MAG), and myelin oligodendrocyte glycoprotein (OMG), RTN4R mediates axonal growth inhibition and may play a role in regulating axonal regeneration and plasticity in the adult CNS. It has been shown that RTN4R performs its inhibitory actions by interacting with the p75 neurotrophin receptor (p75NTR), a TNFRSF member also known for modulating the activities of the Trk family and for inducing apoptosis in neurons and oligodendrocytes. RTN4R may be proposed as a potential drug target for treatment of various neurological conditions such as spinal cord injury, CNS lesions, peripheral nerve injury, stroke and Alzheimer's disease (AD). Additionally, RTN4R may play a role in regulating the function of the gap junctions.
Full Name
reticulon 4 receptor
References

    1. Wang, X. et al., 2006, Ann Neurol. 60(5): 540-549.      

    2. Wang, Y.Z. et al., 2006, Neuroreport.17(6):605-609.       

    3. Zhu, H.Y. et al., 2007, Hum Pathol. 38(3): 426-434.           

    4. David, S. et al., 2008, Trends Neurosci. 31(5): 221-226.       

    5. Jiang, W. et al., 2009, Transl Res. 154(1): 40-48.      

    6. Zhang, L. et al., 2009, J Neurosci, 9(19): 6348-6352.

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