8- (6- Aminohexylamino)adenosine- 3', 5'- cyclic monophosphorothioate, Rp- isomer, immobilized on agarose gel (Rp-8-AHA-cAMPS-Agarose) Biochemical

8- (6- Aminohexylamino)adenosine- 3', 5'- cyclic monophosphorothioate, Rp- isomer, immobilized on agarose gel (Rp-8-AHA-cAMPS-Agarose)

Cat. Num. AAA52690

• Purity: The purity of each ligand to be attached is better than 98% (HPLC).
• Synonyms: 8- (6- Aminohexylamino)adenosine- 3', 5'- cyclic monophosphorothioate, Rp- isomer, immobilized on agarose gel (Rp-8-AHA-cAMPS-Agarose); N/A; 8- (6- Aminohexylamino)adenosine- 3', 5'- cyclic monophosphorothioate, Rp- isomer, immobilized on agarose gel (Rp-8-AHA-cAMPS-Agarose) biochemical

• Purity/Purification: The purity of each ligand to be attached is better than 98% (HPLC).

• Specification: Suspension in 30 mM Na2HP04 buffer (pH 7).
• Ligand density: approx. 5 umol/ml of settled gel.
• Chromatography: After equilibration with about 10 column volumes of starting buffer the affinity column is loaded with the protein solution, e.g. at 50-200 ul/min. In order to elute other nucleotide-dependent proteins unspecifically bound, the column is washed, e.g. with 1 mM 5'-AMP. For elution of E. coli nucleoside diphosphate kinase it is recommended to wash the column with 10 mM ADP/20mM MgCI₂.; Elution of the target protein is performed by a cyclic nucleotide gradient up to 40 mM. For elution of phosphodiesterases it could be advisable to use the highly hydrolysis-resistant Sp-cAMPS or the 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.
• Toxicity and Safety: Please keep in mind that the in vivo properties of these compounds are not sufficiently characterized up to now.; Avoid skin contact or ingestion and allow only trained personnel to work with it.; Our products are designed, developed and sold for research purposes only. They are intended for in vitro and non-human in vivo laboratory applications. Any other use requires approval of health authorities. Not for drug, household or related uses!
• Preparation and Storage: 8-AHA-cAMPS agarose has sufficient stability for chromatography at room temperature and does not need special care during handling or shipment. Nevertheless for longer storing periods the gel should be kept in the refrigerator at 4 - 8°C since desulfurization yielding immobilized cAMP can occur slowly (about 0.1 % week/20°C). Storage buffer should contain 0.1 % sodium azide for prevention of microbial growth.

Structure

Related Product Information for 8- (6- Aminohexylamino)adenosine- 3', 5'- cyclic monophosphorothioate, Rp- isomer, immobilized on agarose gel (Rp-8-AHA-cAMPS-Agarose) biochemical

The PKA inhibitor Rp-cAMPS and the corresponding agonistic Sp-cAMPS are immobilized to agarose via a hexyl spacer at position 8 of the adenine nucleobase.

Properties of Rp-8-AHA-cAMPS Agarose: The ligand of this gel represents an immmobilized form of the cyclic AMP antagonist Rp-cAMPS. Free Rp-cAMPS was found to prevent dissociation of protein kinase A I & II catalytic from regulatory subunits and thus to inhibit the cAMP signal pathway. In addition Rp-cAMPS is not hydrolyzed by mammalian phosphodiesterases. Both properties make this analogue to be an interesting ligand in affinity chromatography, especially when trying to purify protein kinase A holoenzyme, cyclic nucleotide- dependent phosphodiesterases or other cAMP binding proteins. Immobilized Rp-cAMPS can also be used as a stable general ligand with reduced affinity for various nucleotide binding proteins since POE degradation, the first metabolic step necessary in cyclic nucleotide metabolism, is blocked. Immobilized Rp-cAMPS is also available with a short aminoethylamino spacer; for other spacer types, attachment positions or gels please inquire.
If the target protein is absorbed too strongly on common cAMP agaroses, both gels can used as alternative due to the reduced binding affinity of phosphorothioate-modified cyclic nucleotides to most receptors. Also, the column can be useful for purification of cGMP receptors in case the protein sticks too tightly to a corresponding gel with immobilized cGMP.