Phospholamban, phosphorylated (Ser16)

Phospholamban (PLB) is a 52 amino acid phosphoprotein which regulates the calcium pump of cardiac sarcoplasmic reticulum (SR). PLB is an oligomer of five identical subunits each having a cytoplasmic and transmembrane domain. The cytoplasmic domain (residues 1-25) contains the phosphorylation sites and is highly basic and readily cleaved by proteases; whereas the transmembrane domain (aa25-52) is mostly hydrophobic, protease resistant and stabilizes the pentamer. Following adrenergic stimulation of cardiac muscle, PLB is phosphorylated at Ser16 and at Thr17 which is correlated with stimulation of calcium transport activity across the SR membrane and relaxation of cardiac fibers. \n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 0.5-2ug/ml detects phosphorylated phospholamban from a bovine cardiac preparation that was incubated with 10ug PKA for 10 minutes. Bovine cardiac preparation was treated with PKA or untreated. It was resolved by electrophoresis, transferred to nitrocellulose and probed with P4073-51 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit and a chemiluminescence detection system.\nNote: Do not boil cardiac preparation. Boiling may reduce the 25kD pentamer in to tri-, di- or monomers. In addition, phosphorylation may disrupt the pentamer into dimers and monomers. \nOptimal dilutions to be determined by the researcher.\n\nRecommended Secondary Antibodies\nI1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb\nI1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb\nI1904-46Q: IgG, H&L (HRP) Pab Gt xRb\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

P4073-51

Size

100ug

Applications

WB

Hosts

Rabbit

Reactivities

Bov

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Most, P., et al., J. Biol. Chem. 278: 33,809-33,817 (2003). 2. Mayer, E.J., et al., Biochem. Biophys. Research Comm. 267: 40-48 (2000).

Additional Info

Recognizes Phospho-lamban phosphorylated (Ser16) (5-25kD). Additional non-specific bands may be detected at 75 and 100kD in some lysates. Species Crossreactivity: bovine