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Human PAM protein (active)

Human PAM protein (active). GTX00180-pro
Human PAM protein (active). GTX00180-pro
Human PAM protein (active). GTX00180-pro

Cat No. GTX00180-pro

Application Functional Assay
Species Human
Package
10 μg ($299)
APPLICATION

Application Note

Peptidyl-glycine alpha-amidating monooxygenase (PAM) is an enzyme that is required for the biosynthesis of many signaling peptides. This enzyme mainly includes two domains with distinct catalytic activities, a peptidylglycine alpha-hydroxylating monooxygenase (PHM) domain and a peptidyl-alpha- hydroxyglycine alpha-amidating lyase (PAL) domain. These catalytic domains work sequentially to catalyze neuroendocrine peptides to active alpha-amidated products. Besides, Glucosidase Alpha, Acid (GaA) has been identified as an interactor of PAM, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PAM and recombinant human GaA. Briefly, PAM were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GaA-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-PAM pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of PAM and GaA was in a dose dependent manner.

Observed MW

34 kDa.
PROPERTIES

Form

Lyophilized powder

Buffer

Lyophilized from 20 mM Tris (pH 8.0) with 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.01% SKL, 5% Trehalose, Proclin300. Reconstitute with 20 mM Tris and 150 mM NaCl (pH 8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

Storage

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

Region/Sequence

N-terminal His-Tag; Phe21~Cys288 (NP_000910.2)

Expression System

E. coli

Purity

> 95%

Endotoxin

< 1 EU/μg

Conjugation

Unconjugated

Note

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

Synonyms

peptidylglycine alpha-amidating monooxygenase,PAL,PHM

Background

This gene encodes a multifunctional protein. The encoded preproprotein is proteolytically processed to generate the mature enzyme. This enzyme includes two domains with distinct catalytic activities, a peptidylglycine alpha-hydroxylating monooxygenase (PHM) domain and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain. These catalytic domains work sequentially to catalyze the conversion of neuroendocrine peptides to active alpha-amidated products. Alternative splicing results in multiple transcript variants, at least one of which encodes an isoform that is proteolytically processed. [provided by RefSeq, Jan 2016]

Database

Research Area

DATA IMAGES
Human PAM protein (active). GTX00180-pro

GTX00180-pro Functional Assay Image

Functional ELISA analysis of GTX00180-pro Human PAM protein (active) which can bind immobilized GaA protein.

Human PAM protein (active). GTX00180-pro

GTX00180-pro Image

SDS-PAGE analysis of GTX00180-pro Human PAM protein (active).

Human PAM protein (active). GTX00180-pro

GTX00180-pro Image

WB analysis of GTX00180-pro Human PAM protein (active).

REFERENCE
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REVIEW
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