|Detection of Human and Mouse ATN1 by Western Blot. Western blot shows lysates of human liver tissue and mouse liver tissue. PVDF Membrane was probed with 1 µg/mL of Human/Mouse ATN1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6567) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for ATN1 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|ATN1 in Human Brain. ATN1 was detected in immersion fixed paraffin-embedded sections of human brain (hypothalamus) using Sheep Anti-Human/Mouse ATN1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6567) at 3 µg/mL 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 using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
ATN1 (Atrophin-1; also DRPLA protein) is a 190‑200 kDa member of the Atrophin family of proteins that is cleaved into 120‑150 kDa fragments. ATN1 is a ubiquitously expressed transcriptional coactivator. Human ATN1 is 1185 amino acids (aa) in length. It contains multiple motifs, including an NLS (aa 16-32), interspersed poly-Pro, poly-Ser and poly-His regions (aa 376-707), two RE (ArgGlu) repeats (aa 816-934), and an NES (aa 1033-1042). There are at least 17 utilized phosphorylation sites and one acetylated Lys. ATN1 is most characterized by a poly-Gln region between aa 484-497. Normally, there are about 20 consecutive Gln residues, but this number may be increased to more than 70 in pathologic conditions. Proteolytic cleavage generates large C-terminal fragments of 120-150 kDa size. These are unlikely to contain the NLS, and thus are typically cytosolic. ATN1 is suggested to form heterodimers with full-length ATN2/RERE, thus generating a transcriptional repressor. There are multiple potential isoforms. One shows an alternative start site at Met527, while others differ in the number of glutamines in the poly-Gln region. Over aa 1-100, human ATN1 shares 94% aa identity with mouse ATN1.
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