Technical Data
P4073-50
Phospholamban
Description:
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 (residues 25-52) 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.

Applications:
Suitable for use in ELISA, Western Blot, Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-2ug/ml. Detects phospholamban in 20ug bovine cardiac preparation. DO NOT BOIL CARDIAC PREPARATION. Bovine cardiac preparation was resolved by electrophoresis, transferred to nitrocellulose and probed with P4073-50 (1ug/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
Immunohistochemistry: 10ug/ml. Detected phospholamban in frozen rat heart tissue sections, fixed for either 5 minutes with ice cold ethanol:acetic acid [95:5] for 30 minutes with 4% paraformaldehyde, 4% sucrose in PBS, followed by permeabilization with 0.25% Triton X-100 in PBS.
Immunoprecipitation: 4ug/500ug bovine cardiac preparation.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG10.T.110Highly Purified
SizeStorageShippingSourceHost
50ug4C (-20C Glycerol)Blue IceBovineMouse
Concentration:
~0.5mg/ml
Immunogen:
Bovine phospholamban purified from bovine cardiac sarcoplasmic reticulum. Molecular Weight: ~25kD
Purity:
Purified by hydroxyapatite chromatography
Form
Supplied as a liquid in 20mM Tris-HCl, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, before the addition of glycerol to 40%.
Specificity:
Recognizes and is specific for the four forms of bovine phospholamban: phosphorylated and non-phosphorylated phospholamban and the breakdown protein derivatives from the sarcoplasmic reticulum. Species Crossreactivity: canine, rat, mouse and porcine.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Suzuki and Wang, J. Biol. Chem. 261: 7018, 1986. 2. Suzuki, Lui and Wang, Biochem. Cell Biol. 65: 302, 1987. 3. Li, Wang and Coyler, Biochemistry 29: 4535, 1990. Application 4. Slack, J.P., et al., J. Biol. Chem. 272: 1886218868, 1997.