Santa Cruz Biotechnology, Inc.
Neutrophil Elastase Antibody (201)
sc-59335
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Detection of neutrophil elastase (NE) is important in the studies of inflammation, specifically in studying the inflammatory responses of neutrophils. NE is a powerful serine protease that is expressed mainly by neutrophils, which release NE upon stimulation. Specific NE activity is measured by a peptide cleavage assay, but it is also important to know how much of total NE has been released by neutrophils under specific conditions. Total NE could be determined by western blotting in comparison with the known quantities of purified NE. The mouse monoclonal antibody sc-59335 (201) from Santa Cruz was designed to detect human NE by western blotting and other techniques, including IP, IF and ELISA. We tested this antibody for the ability to detect human purified NE in western blotting.
Western Blot
Purified neutrophil elastase, 0.5 microgram; U937 cell lysate, 50 micrograms
2 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; 1:3,000; 1 hr, room temperature
None
West Pico
In our hands, the antibody sc-59335 did not recognize either human NE purified from peripheral blood neutrophils. This antibody also did not produce any a positive NE-related band (should be ~30 kDa) in the U-937 cell lysate (surprisingly used as a positive control for NE by the manufacturing company).
N/A
Nothing noted
This antibody did not recognize human NE
This mAb that was raised against a peptide sequence shared by human NE unfortunately does not recognize purified human NE.