Peptidylglycine Alpha Amidating Monooxygenase (PAM) Active Protein | PAM active protein

Active Peptidylglycine Alpha Amidating Monooxygenase (PAM)

Cat. Num. AAA150334

• Reactivity: Homo sapiens (Human)
• Applications: Activity Assay, Cell Culture
• Purity: >90%
• Synonyms: Peptidylglycine Alpha Amidating Monooxygenase (PAM); N/A; Active Peptidylglycine Alpha Amidating Monooxygenase (PAM); PAM active protein

• Host: E. coli
• Reactivity: Homo sapiens (Human)
• Purity/Purification: >90%
• Form/Format: PBS, pH7.4, containing 0.01% SKL, 5% Trehalose
• Concentration: Original Concentration: 100ug/mL (varies by lot)

• Applicable Applications for PAM active protein: Activity Assay, Cell Culture
• Source: Prokaryotic expression
• Residues: Phe21~Cys288
• Tags: N-terminal His-tag
• Endotoxin Level: <1.0EU per 1ug (determined by the LAL method).
• Predicted isoelectric point: 8.8
• Usage: Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.
• Preparation and Storage: Storage:; Avoid repeated freeze/thaw cycles.; Store at 2-8 degree C for one month. ; Aliquot and store at -80 degree C for 12 months.; ; Stability Test:; The 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 degree 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.

SDS_PAGE

(Sample: Active recombinant PAM, Human)

Bioactivity

(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 alphahydroxylating 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 100uL were then transferred to GaA-coated microtiter wells and incubated for 2h at 37°C. 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 37°C. Finally, add 50uL stop solution to the wells and read at 450nm immediately. The binding activity of PAM and GaA.was shown in Figure 1, and this effect was in a dose dependent manner.)

Gene Sequence

(Gene Sequencing (extract))

Sequence