Analysis Note

ControlBovine cardiac preparation, treated with PKA.

Application

Research Sub CategoryMuscle PhysiologyGeneral Post-translation Modification

This Anti-phospho-Phospholamban (Ser16) Antibody is validated for use in WB for the detection of phospho-Phospholamban (Ser16).

Research CategoryMetabolismSignaling

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General 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 to 25) contains the phosphorylation sites and is highly basic and readily cleaved by proteases; whereas the transmembrane domain (residues 25 to 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.

Immunogen

KLH-conjugated, synthetic peptide corresponding to amino acids 14-25 of human cardiac phospholamban (RA[pS]TIEMPQQAR-C) with an added C-terminal cysteine. The immunizing sequence is identical in mouse, rat, rabbit, pig and dog.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified rabbit polyclonal IgG in buffer containing 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl, 0.07% sodium azide and 30% glycerol. Store at -20°C.

Immunoaffinity Chromatography

Quality

Routinely evaluated by western blot on bovine cardiac preparation that was incubated with PKA.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected phosphorylated phospholamban from a bovine cardiac preparation that was incubated with 10 µg PKA (Catalog # 14-114) for 10 minutes.
DO NOT BOIL CARDIAC PREPARATION.
Boiling may reduce the 25 kDa pentamer into tri-, di- or monomers. In addition, phosphorylation may disrupt the pentamer into dimers and monomers.

Specificity

Recognizes phosphorylated phospho-lamban (Ser16), Mr 5-25 kDa. Additional non-specific bands may be detected, Mr 75 & 100 kDa, in some lysates.

Predicted to cross-react with human, mouse, rat, rabbit, pig and dog.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

Varies