Neuroplastin 65 in Human Brain.
Neuroplastin 65 was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using 3 µg/mL Goat Anti-Human/Mouse Neuroplastin 65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5360) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human/Mouse Neuroplastin 65 by Western Blot.
Western blot shows lysates of human brain and mouse brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Neuroplastin 65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5360) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Neuroplastin 65 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Neuroplastin 65
Neuroplastin (NPTN; also stromal cell-derived receptor 1 and Np55) is a 52-57 kDa member of the Ig-superfamily. It is widely expressed and likely serves as a cell adhesion molecule in multiple tissues. Human Neuroplastin is 282 amino acids (aa) in length. It is a type I transmembrane glycoprotein that contains two Ig-like domains (aa 32-119 and 122-213) and a 38 aa cytoplasmic region (aa 245-282). There is a brain-enriched 65 kDa form (Np65) that negatively regulates LTP in the hippocampus. It contains an N terminal 116 aa Ig-like domain-containing insert after Asn30 and interacts homotypically. Notably, human and rodent Np65 expression patterns are different in brain, with rodent Np65 not predominating in cerebellum. Over aa 29-336, human Np65 is 95% aa identical to mouse Np65.
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