rabbit ngbr antibody  (Proteintech)

Journal: Frontiers in Cellular Neuroscience

Article Title: Schwann Cell Expressed Nogo-B Modulates Axonal Branching of Adult Sensory Neurons Through the Nogo-B Receptor NgBR

doi: 10.3389/fncel.2015.00454

Figure Lengend Snippet: Immunohistochemistry of Nogo-B in the sciatic nerve. (A) The two isoforms Nogo-A and Nogo-B differ considerably in size and function. A common feature is the C-terminal RHD which harbors not only an ER anchoring motif at the very C-terminus but also two hydrophobic sequence stretches (shaded in gray); they are believed to contribute to ER tubulation by inducing membrane curvature. Nogo-A is selectively recognized by the monoclonal 11C7 antibody while the epitope of the Bianca antiserum resides within the NIR domain that is shared by both isoforms. The NEP1–40 peptide is derived from and blocks the signaling activities of the Nogo-66 loop domain. The Nogo-B specific receptor NgBR, and recombinant NgBR-Ecto, bind to the AmNogo-B sequence stretch that is composed of parts of the NIR domain and RHD. (B) Immunolabeling of cross sections of the sciatic nerve of a wildtype and nogo-a/b knockout mouse. The external cytoplasmic compartment of Schwann cells is immunoreactive for Nogo. The myelin sheath that is visualized with myelin basic protein (MBP) is devoid of Nogo. Axons are revealed with a neurofilament antibody. Single confocal sections are shown. Scale bar is 10 μm. (C) Immunolabeling of cross sections of the rat sciatic nerve with the Bianca antiserum and counter-staining with 11C7. Schwann cells are strongly positive for Bianca but not for 11C7 indicating that the majority of Bianca immunoreactivity is due to Nogo-B. In contrast, the Bianca immunoreactivity in axons seems to be due to Nogo-A as demonstrated by the 11C7 signal (white arrows). Scale bar is 5 μm. A single confocal section is shown. (D) Immunolabeling of teased nerve fibers of the sciatic nerve of an adult wildtype mouse. There is prominent Nogo immunoreactivity in Cajal bands and, to a lesser extent, in Schmidt-Lanterman incisures (arrow heads) and transverse trabeculae (triangles) as revealed by MAG and Vimentin co-distribution. The Nogo staining typically extends into nodes of Ranvier (arrows). Single confocal sections are shown. Scale bar is 10 μm.

Article Snippet: For detection the following primary antibodies were used: rabbit Bianca antiserum (1:20,000), rabbit NgBR antibody (1:400; proteintech #15986-1-AP), rabbit PO antibody (1:500; proteintech #10572-1-AP), rabbit p75 NTR antibody (1:1000; Promega #G3231), mouse actin antibody (1:10,000; Chemicon #MAB1501), mouse L-caldesmon (1:500; BD Biosciences #610660).

Techniques: Immunohistochemistry, Sequencing, Membrane, Derivative Assay, Recombinant, Immunolabeling, Knock-Out, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Schwann Cell Expressed Nogo-B Modulates Axonal Branching of Adult Sensory Neurons Through the Nogo-B Receptor NgBR

doi: 10.3389/fncel.2015.00454

Figure Lengend Snippet: Blocking the Nogo-B receptor NgBR mimics the loss-of- nogo branching phenotype. (A) Quantification of morphological parameters of adult wildtype sensory neurons grown on wildtype Schwann cells in the presence or absence of NEP1–40. Five independent experiments were carried out with a total of 181 neurons being analyzed in the control and 177 neurons in the NEP1–40 group. Morphological parameters were normalized to the control (PBS; set to 1.0) and expressed as mean ± SEM. P -values ≤ 0.05 are indicated. Neuronal cell body diameters are depicted in a dot blot. (B) Immunocytochemistry of a Schwann cell neuron co-culture stained for NgBR, Hoechst and neurofilament. Scale bar is 10 μm. (C) Co-immunoprecipitation experiments with a co-culture of wildtype Schwann cells and nogo-a/b deficient sensory neurons. For pull-down Bianca antiserum or a rabbit control IgG was used. Detection was done with Bianca antiserum and an NgBR antibody. Asterisks indicate the rabbit IgG heavy chain that is detected by the secondary anti-rabbit antibody. (D) Quantification of morphological parameters of adult wildtype sensory neurons grown on wildtype Schwann cells in the presence or absence of NgBR-Ecto. Five independent experiments were carried out with a total of 142 neurons being analyzed in the control and 167 neurons in the NgBR-Ecto group. Morphological parameters were normalized to the control (50% glycerol in PBS; set to 1.0) and expressed as mean ± SEM. P -values ≤ 0.05 are indicated. In the presence of NgBR-Ecto TNIS is reduced to 0.75 and TNL to 0.74 relative to the control levels. Neuronal cell body diameters are depicted in a dot blot.

Article Snippet: For detection the following primary antibodies were used: rabbit Bianca antiserum (1:20,000), rabbit NgBR antibody (1:400; proteintech #15986-1-AP), rabbit PO antibody (1:500; proteintech #10572-1-AP), rabbit p75 NTR antibody (1:1000; Promega #G3231), mouse actin antibody (1:10,000; Chemicon #MAB1501), mouse L-caldesmon (1:500; BD Biosciences #610660).

Techniques: Blocking Assay, Control, Dot Blot, Immunocytochemistry, Co-Culture Assay, Staining, Immunoprecipitation