Mouse/Rat Neurocan Antibody

Detection of Mouse/Rat Neurocan by Western Blot.

Western blot shows lysates of rat embryonic hippocampal neurons. PVDF membrane was probed with 1 µg/mL of Mouse/Rat Neurocan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5800) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Neurocan at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Neurocan in Mouse Brain.

Neurocan was detected in perfusion fixed frozen sections of mouse brain (medulla) using Mouse/Rat Neurocan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5800) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Neurocan by Immunohistochemistry

Expression of neurocan and NCAM in postnatal and adult brain. (A) Immunoblot of total brain extracts for neurocan, NCAM, PSA-NCAM, and GAPDH (loading control). (B) Co-localization of tdTomato puncta (red) with the presynaptic marker GAD65 (green) around a neuronal soma (blue). Scale bar = 5 μm. (C) Immunofluorescent staining of neurocan (green) around PV+ tdTomato-labeled interneurons (red) in mouse frontal cortex at P28 and P60. Scale bar = 50 μm. (D) Confocal images of neurocan (green) around perisomatic synaptic puncta (red) onto MATH-2 expressing pyramidal neurons (blue nucleus, outlined with white border). A magnified inset is indicated with a white box. Scale bars = 2.5 μm. (E) Electron micrograph of immunogold labeling for neurocan along the membrane of a neuronal soma (black arrows point to examples). Scale bar = 1 μm. (F) Electron micrograph of an inhibitory axon terminal onto a neuronal soma. Neurocan immunogold labeling is indicated with black arrows, and the axon terminal (AT) is labeled with a white arrowhead. Cytoplasm and nucleus of the soma are identified with text. Scale bar = 1 μm. (G) Verification of neurocan antibody specificity by immunostaining of COS7 cells transfected with Neurocan-AP (+control) or AP (−control), and a no primary antibody control. Scale bar = 50 μm.(H) Immunofluorescent localization of NCAM and neurocan in GABA+ and GABA− cortical neuron cultures. Confocal images of GABA (green), NCAM (blue), and neurocan (red) in cortical neuron cultures. White arrow indicates a GABA-negative neuron positive for NCAM and neurocan, and the green neuron represents a GABAergic interneuron. Scale bar = 10 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29670169), licensed under a CC-BY license. Not internally tested by R&D Systems.