Detection of Human, Mouse, and Rat Inorganic Pyrophosphatase/PPA1 by Western Blot.
Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, Hepa 1‑6 mouse hepatoma cell line, IMR‑32 human neuroblastoma cell line, Neuro‑2A mouse neuroblastoma cell line, and C6 rat glioma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat Inorganic Pyrophosphatase/PPA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6557) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Inorganic Pyrophosphatase/PPA1 at approximately 33 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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: Inorganic Pyrophosphatase/PPA1
PPA1 (Inorganic pyrophosphatase 1; also PPase and IOPPP) is a 32-36 kDa cytoplasmic member of the PPase family of enzymes. It is ubiquitously expressed, and acts on di- (or pyro) phosphate, generating orthophosphate in a Mg+-dependent manner. This activity can both generate energy for cells, or in the case of osteoblasts, provide raw material for calcification. The consumption of pyrophosphate may also remove inhibitors of enzymes such as guanylyl cyclase, and PPA1 itself is also reported to stimulate gene expression. Human PPA1 is 289 amino acids (aa) in length. There is one pyrophosphatase domain between aa 42-255, and two utilized acetylation sites at Lys 57 and Lys228. PPA1 is known to form homodimers. There is one potential alternative start site at Met46. Full-length human PPA1 shares 94% aa sequence identity with mouse PPA1.
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