Neprilysin Activity Assay Kit (Fluorometric), Eukaryotes

Description

Enkephalinase (NEP, EC 3.4.24.11), also known as neutral endopeptidase, enkephalinase, CD10 and common acute lymphoblastic leukemia antigen, is a zinc-containing transmembrane metalloproteinase. It can hydrolyze very important endogenous peptides, such as natriuretic atrial factor, enkephalin, substance P, bradykinin and amyloid beta (ABeta) peptide. Therefore, NEP is a potential therapeutic target in important pathological conditions such as cardiovascular disease, prostate cancer and Alzheimer's disease. NEP has also been used as a biomarker for childhood leukemia, and the detection of NEP in endometrial stromal cells is proposed as a useful tool for diagnosing endometriosis. NEP is currently the main focus of cardiovascular and neurological research. Analysis Wizard's Neprilysin activity kit utilizes the ability of active NEP to cleave synthetic substrates (Abz-based peptides) to release free fluorophores. The released Abz can be easily quantified using a fluorescence microplate reader. The substrate is specific to NEP, which can distinguish the NEP activity in biological samples from trypsin and other structurally similar zinc metalloproteinases, such as angiotensin converting enzymes (ACE1, ACE2), endothelin converting enzymes (ECE1, ECE2) ). Our assay kit is simple and specific, and can detect NEP activity as low as 20 µU/mg.

Species Reactivity

Eukaryote

Application Notes

NEP Activity Assay is highly sensitive with a detection limit of approximately 20 /mg

Relevant Diseases

Neurological Disorders

Detection Method

Fluorometric (Ex/Em = 330/430 nm)

Assay Type

Activity Assay Kit

Components

NEP Assay Buffer
Neprilysin (Lyophilized)
NEP Substrate (in DMSO)
Abz-Standard (1 mM)

Shipping

Gel Pack

Storage

-20°C

Handling Advices

The antibody solution should be gently mixed before use.

Quality Control

Includes NEP positive control

Research Use Only

For research use only

Target

MME

Official Name

MME

Full Name

Membrane metallo-endopeptidase

Alternative Names

MME; Membrane metallo-endopeptidase

Gene ID

4311(Human); 17380(Mouse); 24590(Rat)

Uniprot ID

P08473(Human); Q61391(Mouse); P07861(Rat)

Figure 1 Abz-Standard Curve, results from multiple experiments.


Figure 2 Measurement of purified NEP (10 ng), ACE1 (150 ng), ACE2 (0.5 ng), ECE1 (15 ng) and ECE2 (20 ng) activities using our proprietary substrate. The kit can efficiently distinguish NEP activity from other zinc metalloproteases in biological samples.


Figure 3 Measurement of NEP activity in Rat Lung (10 μg protein) and Rat Kidney (8.5 μg protein). All assays were performed following kit protocol.