Pan-Nogo/Reticulon4

Accession/Synonym: Reticulon 4A, ASY; Nbla00271; Nbla10545; NI220/250; NOGO; NSP; NSP-CL; RTN-X; RTN4-A;  RTN4-B1; RTN4-B2; RTN4-C

The Nogo gene belongs to the family of reticulons (RTN) encoding genes, RTN are associated with the endoplasmic reticulum and are involved in neuroendocrine secretion or in membrane trafficking in neuroendocrine cells.  RTN are highly conserved proteins involved in a variety of physiological functions ranging from gain in function associated with some tissues to apoptosis. They contain 4 RTN family members, RTN (1-4) and are predominantly located in the Endoplasmic Reticulum.  RTN4, also known as NogoA is expressed in several splice variants and give rise to RTN4B/NogoB and RTN4C/NogoC (1).  NogoA, or neurite outgrowth inhibitor, is expressed in white-matter oligodendrocytes, NogoC is expressed in neurons and skeletal muscle, while NogoB is expressed ubiquitously- concentrated in endothelial, smooth muscle and inflammatory cells.  The RTN4 member of the reticulon family-along with other myelin derived neurite outgrowth inhibitor proteins-binds to and interacts with Nogo receptor 1 (NgR1).

CNS injuries often resulted in to irreversible permanent damage since mammalian central nervous system (CNS) has a limited capability of regenerating lost or damaged nerve fibers.  Although CNS axons are capable of regenerating, they do not regenerate or repair due to myelin derived neurite outgrowth inhibitors.  NogoA is a potent inhibitor of axon outgrowth and plasticity.  NogoA also plays a role in regulating axon outgrowth and plasticity during development (2).  NogoA interacts with its receptor Nogo Receptor (NgR) and the paired immunoglobulin like receptor B (PirB) (3). The NogoA binding domain is a 66 amino acids sequence that interacts with NgR and this interaction is mainly responsible for neurite growth inhibition. There are other myelin associated factors such as myelin associated glycoprotein (MAG) and oligdendrocyte associated glycoprotein (OMgp)factors including, Once bound, they serve to limit axonal outgrowth after injury to the CNS preventing neuronal regeneration. Nogo gene is on chromosome 2p16.3 and codes for 1192 aa (NogoA); 986aa (NogoE) and several other smaller transcript variants derived from alternate splicing and differential promotor usage. 

The NogoA/E-selective antibodies were generated against antigenic peptide derived from the C-terminal region of the Nogo gene.  The terminal 23mer peptide antibodies were affinity purified by immobilized antigen based affinity chromatography and stabilized in antibody stabilization buffer for long-term storage. The Nogo antibodies label proteins of apparent MWs of 145k Da; 119kDa and 55 kDa bands in PC-Nogo samples. Nogo-selective antibodies are also available in FITC conjugated preparations for confocal and immunocytochemical analyses.  FabGennix Inc. will also conjugate antibodies to many other fluorescent probes upon request at a nominal charge.  Limited quantities of antigenic blocking peptide and western blot positive controls are also available (inquire before ordering).   FabGennix Inc. has an inventory of more than 1200 well characterized antibodies for research and diagnostic applications, for a complete listing please visit www.FabGennix.com. 

Immunogen:            Synthetic peptide taken from human Nogo AS protein taken from within the C-terminal region corresponding to amino acids 1169-1192 (nkn vkd ama kip aki pgl lkrk ae).  The peptide was conserved in several other species.  The selected peptide was post-synthetically modified to achieve highest antigenicity before coupling to carrier protein using hetero bifunctional cross linker for immunogen preparation.       

Concentration:         Affinity purified Nogo-101AP and Nogo-FITC antibodies (0.59-0.63mg/ml) were stabilized in antibody stabilization buffer for long-term storage.

Applications:           Nogo-101AP antibody was tested in ELISA and western blotting applications at 1:500 dilution using PC-Nogo samples.  Antibody dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions.  Application of this antibody in other protocols has not yet tested. WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d.  Investigators using this antibody in protocols other than listed above can request a complimentary sample of this antibody.                      

Protocols:                Standard protocol for various applications (WB, IMM, IHC) of this antibody can be obtained upon request. The specification sheet for Nogo antibody will be supplied with each product.  FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

                  antibodies for research and diagnostic applications, for a complete listing please visit www.FabGennix.com.