Bioactivity
(Figure 3. The sandard curve of Dns-Tyr-Val-NH2)
Peptidylglycine Alpha Amidating Monooxygenase (PAM) Active Protein | PAM active protein
Active Peptidylglycine Alpha Amidating Monooxygenase (PAM)
Bioactivity
(Figure 3. The sandard curve of Dns-Tyr-Val-NH2)
Bioactivity
(Figure 3. The sandard curve of Dns-Tyr-Val-NH2)
Bioactivity
(Figure 2. The reaction product compared with standard Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2.)
Bioactivity
(Figure 2. The reaction product compared with standard Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2.)
Bioactivity
(Peptidyl-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 was using Dns-Tyr-Val-Gly as substrate, thus the activity of recombinant human PAM was measured by its ability to hydrolyze Dns-Tyr-Val-Gly to Dns-Tyr-Val-NH2. The reaction was performed in 100 mM MES/KOH pH 6.0, 30 mM KI, 30 mM KCl, 1 uM cupric sulfate, 100 ug/ml catalase (APC418Hu05), 1% (v/v) ethanol, 0.001% (v/v) Triton X-100 and 10 mM ascorbate. 250 ul 0.7 mM substrate of Dns-Tyr-Val-Gly was added with 250 ul various concentrations of recombinant human PAM (1 ug/ml,5 ug/ml, 10 ug/ml and 20 ug/ml). Incubated at 37 degree C for 30min, the reaction was stopped by addition 6% (v/v) TCA. The product and substrate was detected by RP-HPLC with UV-detection at 280 nm, the analyses were performed at 25 degree C employing a Agilent ZORBAX Poroshell SB C18 column (9.4×250 mm, 5 um), the flow rate was 1 ml/min. The mobile phase consisted of 100 mM sodium acetate (pH 6.5) and 30 min linear gradient of 10-90% acetonitrile. At 30-35 min, the mobile phase consisted of 90% acetonitrile and 10% sodium acetate. The result was shown in Figure 1. As the Figure 1 shows, the substrate have been hydrolyzed to Dns-Tyr-Val-NH2 after incubating with recombinant human PAM. The retention time of Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2 is 24.088 and 30.421 respectively (Figure 2). The specific activity of recombinant human PAM is >6600 pmol/min/ug.)
Bioactivity
(Peptidyl-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 was using Dns-Tyr-Val-Gly as substrate, thus the activity of recombinant human PAM was measured by its ability to hydrolyze Dns-Tyr-Val-Gly to Dns-Tyr-Val-NH2. The reaction was performed in 100 mM MES/KOH pH 6.0, 30 mM KI, 30 mM KCl, 1 uM cupric sulfate, 100 ug/ml catalase (APC418Hu05), 1% (v/v) ethanol, 0.001% (v/v) Triton X-100 and 10 mM ascorbate. 250 ul 0.7 mM substrate of Dns-Tyr-Val-Gly was added with 250 ul various concentrations of recombinant human PAM (1 ug/ml,5 ug/ml, 10 ug/ml and 20 ug/ml). Incubated at 37 degree C for 30min, the reaction was stopped by addition 6% (v/v) TCA. The product and substrate was detected by RP-HPLC with UV-detection at 280 nm, the analyses were performed at 25 degree C employing a Agilent ZORBAX Poroshell SB C18 column (9.4×250 mm, 5 um), the flow rate was 1 ml/min. The mobile phase consisted of 100 mM sodium acetate (pH 6.5) and 30 min linear gradient of 10-90% acetonitrile. At 30-35 min, the mobile phase consisted of 90% acetonitrile and 10% sodium acetate. The result was shown in Figure 1. As the Figure 1 shows, the substrate have been hydrolyzed to Dns-Tyr-Val-NH2 after incubating with recombinant human PAM. The retention time of Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2 is 24.088 and 30.421 respectively (Figure 2). The specific activity of recombinant human PAM is >6600 pmol/min/ug.)
SDS-PAGE
(Figure. SDS-PAGE)
SDS-PAGE
(Figure. SDS-PAGE)
NCBI and Uniprot Product Information
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Product Notes
The PAM pam (Catalog #AAA161894) is an Active Protein produced from 293F cell and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Peptidylglycine Alpha Amidating Monooxygenase (PAM) can be used in a range of immunoassay formats including, but not limited to, Activity Assay, Cell Culture. Researchers should empirically determine the suitability of the PAM pam for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Phe31~Val8 17. It is sometimes possible for the material contained within the vial of "Peptidylglycine Alpha Amidating Monooxygenase (PAM), Active Protein" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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