Detection of Human ORP150/HSP12A by Western Blot.
Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and human breast cancer tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human ORP150/HSP12A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5558) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for ORP150/HSP12A at approximately 150 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.
ORP150 (Oxygen-related protein of 150 kDa) is a member of the heat shock protein 70 family of molecules. Although the predicted MW is 111 kDa, it runs at 150 kDa in SDS-PAGE. The discrepancy may not be anomalous, since it is secreted into the ER lumen and likely to be glycosylated and/or phosphorylated. ORP150 is highly expressed in cells involved in secretion such as pancreatic acinar cells and hepatocytes. Mature human ORP150 is 967 amino acids (aa) in length. It contains two HSP70 signatures (aa 230‑243 and 380‑394) and an ER-retention signal (aa 996‑999). There are multiple isoforms. Three show alternate start sites at Met88, Met152, and a Met 44 aa upstream of the standard start site. Four isoforms also show deletions of aa 148‑958, 51‑59, 603‑664 and 164‑507, respectively. There is one 44 aa substitution for aa 603‑999. Over aa 1‑994, human ORP150 shares 92% aa identity with mouse ORP150.
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