Santa Cruz Biotechnology
Neutrophil Elastase Antibody
sc-55549
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The detection of human neutrophil elastase (NE) is important in enzymology, where NE release from neutrophils or its presence should be confirmed and quantified.The antibody sc-55549 was designed to detect human neutrophil elastase in western blotting and immunofluorescence. We used this antibody to compare in the same samples the total amount of NE released from human neutrophils versus NE activity (determined by substrate cleavage assay).
Western Blot
Purified neutrophil elastase, 1 microgram; U937 cell lsate, 20 micrograms; human neutrophils, 2x10e^5 cells
1 ug/ml in PBS supplemented with 5% nonfat milk and 0.05% Tween 20; overnight, cold room
5% nonfat milk in PBS/0.05% Tween 20
goat anti-mouse IgG conjugated with HRP; 1:1,000; 1 hr, room temperature
None
West pico
In our hands, this antibody did not recognize neutrophil elastase from any tested sources, including neutrophil elastase purified from human neutrophils loaded up to 1 microgram per lane, contents of 2x10e^5 human neutrophils, and cell lysate of U937 cells, which according to the company should provide positive control. The antibody failed to recognize neutrophil elastase (~29.5 kDa) under both reducing and non-reducing conditions. The image shows negative results under non-reducing conditions.
N/A
The antibody failed to reliably detect human neutrophil elastase in WB.
I would not recommend this antibody for detection of human neutrophil elastase by western blotting