{"product_id":"new-product-619783","title":"Active Peptidylglycine Alpha Amidating Monooxygenase (PAM)","description":"\u003cp\u003eProduct No.\u003c\/p\u003e\n\u003cp\u003eAPC744Ra61\u003c\/p\u003e\n\u003cp\u003eOrganism Species\u003c\/p\u003e\n\u003cp\u003eRattus norvegicus (Rat).\u003c\/p\u003e\n\u003cp\u003eBuffer Formulation\u003c\/p\u003e\n\u003cp\u003ePBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.\u003c\/p\u003e\n\u003cp\u003eTraits\u003c\/p\u003e\n\u003cp\u003eFreeze-dried powder\u003c\/p\u003e\n\u003cp\u003ePurity\u003c\/p\u003e\n\u003cp\u003e\u0026gt; 95%\u003c\/p\u003e\n\u003cp\u003eIsoelectric Point\u003c\/p\u003e\n\u003cp\u003e5.7\u003c\/p\u003e\n\u003cp\u003eApplications\u003c\/p\u003e\n\u003cp\u003eCell culture; Activity Assays.\u003c\/p\u003e\n\u003cp\u003eUOM\u003c\/p\u003e\n\u003cp\u003e10µg 50µg 200µg 1mg 5mg\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cdiv class=\"product\"\u003e\n\u003cdiv class=\"product_left\"\u003e\n\u003ch3\u003eACTIVITY TEST\u003c\/h3\u003e\n\u003cdiv\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cdiv class=\"product_image\"\u003e\n\u003cimg src=\"https:\/\/www.cloud-clone.com\/upload\/product\/pictures\/APC744Ra61-201803260926482475.jpg\"\u003e\n\u003cp\u003eFigure. Recombinant Rat PAM activity assay by HPLC.\u003cbr\u003ePeptidyl-glycine alpha-amidating monooxygenase (PAM) is an enzyme that is required for the biosynthesis of many signaling peptides. It has two enzymatically active domains with catalytic activities-peptidylglycine alpha-hydroxylating monooxygenase (PHM) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL). These catalytic domains work sequentially to catalyze neuroendocrine peptides to active alpha-amidated products. A typical activity assay using Dns-Tyr-Val-Gly as substrate, thus the recombinant rat PAM activity was measured by its ability to hydrolyze Dns-Tyr-Val-Gly to Dns-Tyr-Val-NH2. The reaction was preformed in 1mL containing 100mM MES\/KOH pH 6.0, 30mM KI, 30mM KCl, 1μmol\/L cupric sulfate, 100ug\/mL catalase, 1% (v\/v) ethanol, 0.001% (v\/v) Triton X-100, 10mM ascorbate, 0.35mM\/L Dns-Tyr-Val-Gly (0.2mg\/mL) and initiated by addition various concentrations of PAM (0.1ug\/mL, 1ug\/mL, 5ug\/mL). Incubated at 37℃ for 30min, the reaction stopped by addition 6% (v\/v) TFA. The product and substrate was detected by RP-HPLC with UV-detection at 280nm, the analyses were performed at 25℃ employing a Agilent ZORBAX Poroshell SB C18 column (9.4×250mm, 5μm), the flow rate was 1ml\/min. The mobile phase consisited of 100 mM sodium acetate (pH 6.5) and 35min linear gradient of 10-90% acetonitrile. The result was shown in Figure 1.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"product_image\"\u003e\n\u003cimg src=\"https:\/\/www.cloud-clone.com\/upload\/product\/pictures\/APC744Ra61-201803260927357451.png\"\u003e\n\u003cp\u003eFigure. The reaction product compared with standard Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2.\u003cbr\u003eAs the Figure 1 shows, after 30min later ,the substrate have been hydrolyzed when the PAM was 5ug\/mL. The retention time of Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2 is 24.315 and 30.806 respectively.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"product_image\"\u003e\n\u003cimg src=\"https:\/\/www.cloud-clone.com\/upload\/product\/pictures\/APC744Ra61-201803260928052023.jpg\"\u003e\n\u003cp\u003eFigure. The sandard curve of Dns-Tyr-Val-NH2.\u003cbr\u003eOne unit of enzyme activity is defined as the amount of enzyme required to convert 1μmol of substrate to amidated product in 1min at 37°C. Thus the recombinant rat PAM activity is 2.4×106U.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch3\u003eUSAGE\u003c\/h3\u003e\n\u003cdiv\u003e\n\u003cp\u003eReconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg\/mL. Do not vortex.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch3\u003eSTORAGE\u003c\/h3\u003e\n\u003cdiv\u003e\n\u003cp\u003eAvoid repeated freeze\/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch3\u003eSTABILITY\u003c\/h3\u003e\n\u003cdiv\u003e\n\u003cp\u003eThe thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cdiv\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"product_relation\"\u003e\n\u003ctable\u003e\n\u003ctbody\u003e\n\u003ctr class=\"first_tr\"\u003e\n\u003ctd class=\"td_1\"\u003eCatalog No.\u003c\/td\u003e\n\u003ctd class=\"td_2\"\u003eRelated products for research use of Rattus norvegicus (Rat) Organism species\u003c\/td\u003e\n\u003ctd class=\"td_3\"\u003eApplications (RESEARCH USE ONLY!)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eEPC744Ra61\u003c\/td\u003e\n\u003ctd\u003e\u003ca href=\"https:\/\/www.cloud-clone.com\/products\/EPC744Ra61.html\"\u003eEukaryotic Peptidylglycine Alpha Amidating Monooxygenase (PAM)\u003c\/a\u003e\u003c\/td\u003e\n\u003ctd\u003ePositive Control; Immunogen; SDS-PAGE; WB.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eAPC744Ra61\u003c\/td\u003e\n\u003ctd\u003e\u003ca href=\"https:\/\/www.cloud-clone.com\/products\/APC744Ra61.html\"\u003eActive Peptidylglycine Alpha Amidating Monooxygenase (PAM)\u003c\/a\u003e\u003c\/td\u003e\n\u003ctd\u003eCell culture; Activity Assays.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv id=\"more_info\"\u003e\u003c\/div\u003e","brand":"Cloud-clone","offers":[{"title":"50ug \/ \u003e 95% \/ Rattus norvegicus (Rat)","offer_id":47569023795480,"sku":"APC744Ra61","price":975.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/5652\/1400\/products\/NP_eb1c926a-b5e8-4d53-ac8f-466e187ca016.jpg?v=1705980908","url":"https:\/\/danabiosci.com\/products\/new-product-619783","provider":"Dana Bioscience","version":"1.0","type":"link"}