{"product_id":"recombinant-human-c-amp-dependant-protein-kinase-a-catalytic-subunit-alpha-bhp11001682","title":"Recombinant Human c-AMP dependant Protein Kinase A catalytic subunit alpha","description":"\u003cp\u003e\u003cstrong\u003eRecombinant Human c-AMP dependant Protein Kinase A catalytic subunit alpha\u003c\/strong\u003e is supplied as a recombinant protein for in vitro research use.\u003c\/p\u003e\n\u003ch3\u003eProduct format\u003c\/h3\u003e\n\u003cp\u003eProvided as a recombinant protein suitable for in vitro workflows such as binding studies, screening, and assay development. Refer to the specifications table for expression format and molecular properties.\u003c\/p\u003e\n\u003ch3\u003eActivity \u0026amp; assay information\u003c\/h3\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cstrong\u003eUnit Definition:\u003c\/strong\u003e One unit is defined as the amount of cAMP-Dependent Protein Kinase, recombinant C? catalytic subunit, required to incorporate 1 pmol of phosphate into the specific substrate peptide kemptide (LRRASLG) in one minute at 30°C.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eSpecific Activity:\u003c\/strong\u003e The specific activity of the recombinant PKA catalytic subunit alpha, is \u0026gt;10,000,000 U\/mg.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eAssay Conditions:\u003c\/strong\u003e Roskoski-AssayProtein kinase activity can be measured using a modified radioactive assay according to Roskoski et al. The assay will be performed in a mixture containing 50mM MOPS (pH7.0), 10mM MgCI2, 0.25 mg\/ml bovine serum albumin, 100 IJM Kemptide (peptide substrate), 100 IJM unlabeled ATP mixed with [y_32p] ATP (500-1000 cpm\/pmol) and Ca subunit in a final volume of 50 IJI. Reaction is started by addition of the Ca subunit and can be stopped after 5 minutes incubation at 30°C by spotting the reaction mix onto Whatman P-81 filters and soaking the filters four times in 75mM phosphoric acid (10 ml per sample) for at least 5 minutes. After four washing steps rinse filters with ethanol, dry and count. Roskoski, R., Jr. (1983) Methods Enzymol. 99, 3-6 For the detection of phosphorylation in substrate proteins the phosphotransferase reaction can alternatively be stopped by taking aliquots of the mixture and adding SDS sample buffer. The phosphorylation status of the substrate proteins can subsequently be analysed using SDS PAGE and autoradiography. Zimmermann, B. (1999) Journal of Biological Chemistry.274, 9, 5370-78.\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"ProSpec-Tany TechnoGene Ltd","offers":[{"title":"1 ug","offer_id":53038068597101,"sku":"pka-200-1UG","price":60.0,"currency_code":"USD","in_stock":true},{"title":"5 ug","offer_id":53038319272301,"sku":"pka-200-5UG","price":145.0,"currency_code":"USD","in_stock":true},{"title":"100 ug","offer_id":53038319305069,"sku":"pka-200-100UG","price":2150.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/Prospecbio-prkaca-PKA-200.png?v=1782158165","url":"https:\/\/www.ebiohippo.com\/products\/recombinant-human-c-amp-dependant-protein-kinase-a-catalytic-subunit-alpha-bhp11001682","provider":"BioHippo","version":"1.0","type":"link"}