The PDE-resistant protein kinase G (PKG) activator Sp-cGMPS immobilized on agarose by an aminohexylcarbamoyl spacer attached to the ribose 2' position of the ligand. The gel can be used for affinity chromatography of various cyclic nucleotide-responsive proteins, especially those which tolerate modification of the 2' ribose hydroxyl group, such as some phosphodiesterases. This type of gel is also available with immobilized PDE-stable Rp-cGMPS (Rp-2'-AHC-cGMPS-Agarose (Prod. No. BLG-A051
)) and with normal cGMP (2'-AHC-cGMP-Agarose (Prod. No. BLG-A059
|Short Term Storage:||Ambient|
|Long Term Storage:||+4°C|
|Miscellaneous/General:||Chromatography: After equilibration with about 10 column volumes of starting buffer the affinity column is loaded with the protein solution at 50 - 100 µl/min. In order to elute other nucleotide-dependent proteins unspecifically bound, the column is washed, e.g. with 1mM 5'- GMP.|
Elution of the target protein is performed by a cyclic nucleotide gradient. For elution of phosphodiesterases it could be advisable to use the highly hydrolysis-resistant phosphorothioate-modified analogs or PDE inhibitors.
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 cGMP 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|
General Literature References
Binding of adenosine 3',5'-monophosphate dependent protein kinase regulatory subunit to immobilized cyclic nucleotide derivatives
: W.L. Dills, Jr., et al.; Biochemistry 15
, 3724 (1976), Abstract
Purification of rabbit skeletal muscle protein kinase regulatory subunit using cyclic adenosine-3':5'-monophosphate affinity chromatography
: W.L. Dills, Jr., et al.; BBRC 62
, 70 (1975), Abstract