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Active Neutrophil Elastase (NE)

ELANE; HLE; HNE; ELA2; PMN-E; Neutrophil Elastase; Medullasin; Polymorphonuclear Leukocyte Elastase; Bone Marrow Serine Protease; Leukocyte Elastase; PMN Elastase

ACTIVITY TEST

Neutrophil elastase (NE), also known as polymorphonuclear leukocyte elastase, is a major protease in the primary granules of neutrophils, is involved in microbicidal activity. It is located primarily in the azurophil granules of polymorphonuclear leukocytes. NE is an important factor promoting inflammation, has bactericidal effects, and shortens the inflammatory process. NE also regulates tumor growth by promoting metastasis and tumor microenvironment remodeling. However, NE plays a role in killing tumors under certain conditions and promotes other diseases such as pulmonary ventilation dysfunction. Additionally, it plays a complex role in various physiological processes and mediates several diseases. Lactoferrin (LTF) has been identified as an interactor of NE, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant rat NE and recombinant goat LTF. Briefly, biotin-linked NE were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to LTF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µl stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant rat NE and recombinant goat LTF was shown in Figure 1, the EC50 for this effect is 1.19 ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

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°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.

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Magazine Citations
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