Detection of Human and Mouse Proprotein Convertase 2/PCSK2 by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line, beta TC‑6 mouse beta cell insulinoma cell line, and HepG2 human hepatocellular carcinoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human Proprotein Convertase 2/PCSK2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6018) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Proprotein Convertase 2/PCSK2 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Proprotein Convertase 2/PCSK2 in HepG2 Human Cell Line.
Proprotein Convertase 2/PCSK2 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human/Mouse Proprotein Convertase 2/PCSK2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue).This application has not been tested in mouse samples. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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: Proprotein Convertase 2/PCSK2
Proprotein Convertase 2 (PCSK2) is a subtilisin-like serine peptidase that processes proteins into biologically active products in the secretory pathway. Like other furin/kexin family members, PCSK2 cleaves its substrates primarily after sites of paired basic amino acid residues (1). Autoactivation of PCSK2 occurs in a post‑Golgi compartment of the secretory system (2). Interaction with the secretory protein 7B2 is required for proper activation of PCSK2 (3). The N-terminal domain of 7B2 stabilizes active PCSK2, and a C-terminal fragment of 7B2 can inhibit PCSK2 activity (4). rhPCSK2 was coexpressed with 7B2 to facilitate activation of the enzyme. PCSK2 is a major proteolytic processing enzyme of the regulated secretory pathway of the neuroendocrine system, where it generates a number of hormones and neuropeptides (5). Products of PCSK2 processing include enkephalins, insulin, somatostatin, dynorphin, and LHRH. Within the region used as an immunogen, human PCSK2 shares 97% aa identity with mouse PCSK2.
Zhou, A. et al. (1999) J. Biol. Chem. 274:20745.
Lamango, N.S. et al. (1999) Arch. Biochem. Biophys. 362:275.
Zhu, X. and I. Lindberg (1995) J. Cell Biol. 129:1641.
Lamango, N. et al. (1996) Arch. Biochem. Biophys. 330:238.
Rouillé, Y. et al. (1995) Front. Neuroendocrinol. 16:322.
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