polyclonal sheep  (R&D Systems)

Journal: BMC Neurology

Article Title: NG2 and phosphacan are present in the astroglial scar after human traumatic spinal cord injury

doi: 10.1186/1471-2377-9-32

Figure Lengend Snippet: Normal distribution of NG2, neurocan, versican and phosphacan in the human spinal cord . Transverse sections of control human spinal cords. A: NG2 immunohistochemistry reveals small stellate-shaped cells distributed homogeneously in white matter regions of human spinal cord (arrows). B: In the white matter, neurocan immunoreactivity is observed in the wall of a small blood vessel (arrow). Furthermore, a reticular staining pattern can be seen. C: In a dorsal nerve root, neurocan staining is present in myelin sheaths. D: Versican immunoreactivity is scattered in a dorsal nerve root and can be found in myelin sheaths of small diameter axons. E: Phosphacan immunohistochemistry reveals a fine reticular staining pattern in the gray matter. F: In a dorsal nerve root, phosphacan-immunopositive myelin rings can be observed. ( A-F magnification × 320).

Article Snippet: The primary antibodies used were: monoclonal mouse anti-human NG2 (clone B5, undiluted cell culture supernatant from ATCC cultures and antibody 9.2.27, diluted 1:50; gift from Prof. R. Reisfeld, Scripps Research Institute), monoclonal mouse anti-neurocan (Chemicon, diluted 1:100), polyclonal rabbit anti-versican (Acris antibodies, diluted 1:1.000), monoclonal mouse anti-phosphacan (Chemicon, diluted 1:500), polyclonal rabbit anti-GFAP (DAKO, diluted 1:2.500), polyclonal rabbit anti-myelin basic protein (MBP) (Chemicon, diluted 1:1.000) and polyclonal rabbit anti-neurofilament (NF, Sigma-Aldrich, diluted 1:2.000).

Techniques: Immunohistochemistry, Staining

Journal: BMC Neurology

Article Title: NG2 and phosphacan are present in the astroglial scar after human traumatic spinal cord injury

doi: 10.1186/1471-2377-9-32

Figure Lengend Snippet: The cellular and molecular composition of the scar in human SCI after early and long survival times . Transverse sections of the human spinal cord of control cases and at early and late survival times after SCI. The schematic diagrams in the upper right corner indicate the region from where the actual picture was taken (black rectangle). A: Co-localisation of phosphacan (green) and MBP (red) confirms the presence of the CSPG in the myelin sheaths of control dorsal root axons. B: Double immunofluorescence with phosphacan (green) and laminin (red) reveals CSPG-immunopositive myelin rings surrounded by laminin-positive endoneurium in a dorsal nerve root of control spinal cord. C: Double immunofluorescence with NG2 (red) and CD68 (green) in macrophages at the lesion epicentre, 10 days after injury. D: One year after SCI, staining for versican (red) and NF (green) in sections from the lesion epicentre demonstrated individual and bundled regenerated nerve fibres that were surrounded by a versican-positive endoneurium in the ECM. E-G: In sections from the lesion epicentre of the same case, double immunofluorescence for NF (green) and neurocan ( E , red), versican ( F , red) or phosphacan ( G , red) revealed nerve fibres surrounded by a CSPG-positive endoneurium. H-I: In the intermediate zone of the same case, GFAP (green) and NG2 ( H , red) and phosphacan ( I , red) immunofluorescence demonstrated the close overlap of all three proteins in the astroglial scar after human SCI. J: In an adjacent section, NF (green) and phosphacan immunohistochemistry revealed occasional, small, nerve fibres still present within the CSPG-rich ECM of the astroglial scar. ( A-J magnification × 400).

Article Snippet: The primary antibodies used were: monoclonal mouse anti-human NG2 (clone B5, undiluted cell culture supernatant from ATCC cultures and antibody 9.2.27, diluted 1:50; gift from Prof. R. Reisfeld, Scripps Research Institute), monoclonal mouse anti-neurocan (Chemicon, diluted 1:100), polyclonal rabbit anti-versican (Acris antibodies, diluted 1:1.000), monoclonal mouse anti-phosphacan (Chemicon, diluted 1:500), polyclonal rabbit anti-GFAP (DAKO, diluted 1:2.500), polyclonal rabbit anti-myelin basic protein (MBP) (Chemicon, diluted 1:1.000) and polyclonal rabbit anti-neurofilament (NF, Sigma-Aldrich, diluted 1:2.000).

Techniques: Immunofluorescence, Staining, Immunohistochemistry

Journal: BMC Neurology

Article Title: NG2 and phosphacan are present in the astroglial scar after human traumatic spinal cord injury

doi: 10.1186/1471-2377-9-32

Figure Lengend Snippet: The cellular and molecular composition of the scar in human SCI after long survival times . Transverse sections of the lesioned human spinal cord at 1 year (A-C) and 20 years (D-I) after injury. The schematic diagrams in the upper right corner indicate the region from where the actual picture was taken (black rectangle). A: NG2 staining revealed a network of irregular lamellae in the ECM at the lesion epicentre. B-C: In near adjacent sections, versican ( B ) and phosphacan ( C ) immunohistochemistry demonstrated fibre-like structures either singly or in small bundles. D: Twenty years after SCI, nerve root-like structures at the lesion epicentre demonstrated neurocan-positive myelin rings. E: In a near adjacent section, phosphacan immunoreactivity was also present in myelin sheaths surrounding axons in neuromas. F: Versican immunopositive myelin rings surrounding regenerated nerve fibres were more scattered and of a small diameter in the nerve root-like structures at the lesion epicentre. G: Twenty years after SCI, diffuse but elevated levels of NG2 immunoreactivity were still associated with the ECM of the connective tissue scar (lower part of image) as well as with the astroglial scar of the intermediate zone (upper part of image). At the lesion epicentre, NG2 staining was located on more loosely arranged sheets of basal lamina-like ECM. In the astrocytic scar, NG2 was associated with a dense irregular network of processes. Arrows demarcate the border between the connective tissue component of the lesion and the adjacent astrocytic scar. H: In a near adjacent section, diffuse phosphacan immunoreactivity was also associated with the dense network of processes of the astroglial scar. No immunoreactivity was associated with the ECM of the connective tissue scar. I: GFAP staining strictly delineated the interface between the astroglial scar and the connective tissue scar. ( A-I magnification × 320).

Article Snippet: The primary antibodies used were: monoclonal mouse anti-human NG2 (clone B5, undiluted cell culture supernatant from ATCC cultures and antibody 9.2.27, diluted 1:50; gift from Prof. R. Reisfeld, Scripps Research Institute), monoclonal mouse anti-neurocan (Chemicon, diluted 1:100), polyclonal rabbit anti-versican (Acris antibodies, diluted 1:1.000), monoclonal mouse anti-phosphacan (Chemicon, diluted 1:500), polyclonal rabbit anti-GFAP (DAKO, diluted 1:2.500), polyclonal rabbit anti-myelin basic protein (MBP) (Chemicon, diluted 1:1.000) and polyclonal rabbit anti-neurofilament (NF, Sigma-Aldrich, diluted 1:2.000).

Techniques: Staining, Immunohistochemistry