The protein kinase A activator Sp-cAMPS immobilized on agarose by an aminohexylamino spacer attached to position 8 of the ligand. Free Sp-cAMPS is only very slowly metabolized by corresponding phosphodiesterases and thus represents an extraordinary stable ligand. In addition, its reduced binding affinity towards protein kinase A compared to cAMP should result in milder desorption conditions in affinity chromatography. The gel can be used for affinity chromatography of various cyclic nucleotide-responsive proteins such as protein kinases, phosphodiesterases and others.
|Formulation:||Liquid. 0.05M Na2HPO4 containing 0.1% sodium azide.|
|Short Term Storage:||Ambient|
|Long Term Storage:||+4°C|
|Miscellaneous/General:||After equilibration with about 10 column volumes of starting buffer the affinity column is loaded with the protein solution at 50-100µl/min. In order to elute unspecifically bound nucleotide-dependent proteins the column can be washed, e.g. with 1mM 5'-AMP. Elution of the target protein is performed by a cyclic nucleotide gradient up to 1mM. For elution of phosphodiesterases it could be advisable to use the highly resistant Sp-cAMPS or the completely stable Rp-cAMPS. For protein kinase holoenzyme elution the gradient should be made with Rp-cAMPS.|
Suitable buffer systems for your special application have to be tested but phosphate should be not optimal since one essential affinity interaction of cyclic nucleotides towards their target receptors is the cyclic phosphate.
Regeneration can be achieved by a combination of up to 100 mM cAMP and buffer salts or 8 M urea.
|Background / Technical Information:||For the Original Manufacturer's Data Sheet please click here.|
Our product description may differ slightly from the Original Manufacturer's Data Sheet.
|Manufacturer:||manufactured by Biolog Life Science Institute|