Rat PAM protein, His tag (active)

Catalogue Number: GTX00374-PRO-GTX

Manufacturer:	GeneTex
Preservative:	ProClin™ 300
Physical state:	Lyophilized
Type:	Bioactive Proteins
Alias:	peptidylglycine alpha-amidating monooxygenase , PHM
Host Cell:	HEK293 cells
Shipping Condition:	Blue Ice
Unit(s):	10 ug
Application:	FA

Overview
Datasheets

Description

Description: catalyzes conversion of glycine-extended peptides into biologically active alpha-amidated peptides [RGD, Feb 2006]

Additional Text

Gene Name

PAM

Protein Tag

HIS Tag

Gene ID

25508

Uniprot ID

P14925

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.

Application Notes

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 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 100 mM MES/KOH (pH 6.0), 30 mM KI, 30 mM KCl, 1µmol/L cupric sulfate, 100 µg/ml catalase, 1% (v/v) ethanol, 0.001% (v/v) Triton X-100, 10 mM ascorbate, 0.35 mM/L Dns-Tyr-Val-Gly (0.2 mg/ml) and initiated by addition various concentrations of PAM (0.1 µg/ml, 1 µg/ml, 5 µg/ml). Incubated at 37C 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 25C employing a Agilent ZORBAX Poroshell SB C18 column (9.4x250mm, 5µm), the flow rate was 1 ml/min. The mobile phase consisited of 100 mM sodium acetate (pH 6.5) and 35min linear gradient of 10-90% acetonitrile. The reaction product compared with standard Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2. After 30min later, the substrate have been hydrolyzed when the PAM was 5µg/ml. The retention time of Dns-Tyr-Val-Gly and Dns-Tyr-Val-NH2 is 24.315 and 30.806 respectively.

Accession Number

NP_037132.2

Storage Note

For short-term storage (1-2 weeks), store at 4C. For long-term storage, store at -20C or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.

GTX00374-PRO Datasheet

Price

£371.00

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