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Anti-Neutrophil Elastase ELANE Rabbit Monoclonal Antibody

All lanes use the Antibody at 1:5K dilution for 1 hour at room temperature.
Figure 1. Western blot analysis of Neutrophil Elastase using anti-Neutrophil Elastase antibody (M01607).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human THP-1 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Neutrophil Elastase antigen affinity purified monoclonal antibody (Catalog # M01607) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Neutrophil Elastase at approximately 29 kDa. The expected band size for Neutrophil Elastase is at 29 kDa.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	P08246
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Human
Clonality	Monoclonal
Format	Liquid
Description	Anti-Neutrophil Elastase ELANE Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF, Flow Cytometry applications. This antibody reacts with Human.

Additional Information
Gene ID	1991
Other Names	Neutrophil elastase, 3.4.21.37, Bone marrow serine protease, Elastase-2, Human leukocyte elastase, HLE, Medullasin, PMN elastase, ELANE, ELA2
Calculated MW	28518 Da
Application Details	WB 1:500-1:2000 IHC 1:100-1:500 ICC/IF 1:50-1:200 FC 1:30
Tissue Specificity	Bone marrow cells.
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: AOCI-5
Immunogen	A synthesized peptide derived from human Neutrophil Elastase
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	ELANE
Synonyms	ELA2
Function	Serine protease that modifies the functions of natural killer cells, monocytes and granulocytes (PubMed:15140022). Shows microbicidal activity in neutrophils by mediating cleavage and degradation of bacterial proteins (PubMed:10947984, PubMed:25161283). Capable of killing E.coli but not S.aureus in vitro; digests outer membrane protein A (ompA) in E.coli and K.pneumoniae (PubMed:10947984). Involved for the formation of neutrophil extracellular traps (NETs): translocates into the nucleus downstream of MPO and mediates degadation of histone H4, thereby participating to chromatin decondensation (PubMed:20974816, PubMed:25066128, PubMed:28574339). NETs are mainly composed of DNA fibers and are released by neutrophils to trap pathogens during inflammation (PubMed:20974816, PubMed:25066128, PubMed:28574339). Inhibits C5a-dependent neutrophil enzyme release and chemotaxis (PubMed:15140022). Promotes cleavage of GSDMB, thereby inhibiting pyroptosis (PubMed:36899106). Promotes blood coagulation (PubMed:20676107). Through the activation of the platelet fibrinogen receptor integrin alpha-IIb/beta-3, potentiates platelet aggregation induced by a threshold concentration of cathepsin G (CTSG) (PubMed:25211214, PubMed:9111081). Cleaves and thus inactivates tissue factor pathway inhibitor (TFPI) (PubMed:20676107, PubMed:25211214).
Cellular Location	Cytoplasmic vesicle, phagosome. Lysosome Cytolytic granule. Nucleus. Note=Localizes to azurophil granules, a type of primary lysosome found in neutrophils and monocytes that contains antimicrobial proteins (PubMed:20974816, PubMed:25066128) Translocates from azurophil granules to the nucleus downstream of MPO during formation of neutrophil extracellular traps (NETs), and is present in released NETs (PubMed:20974816, PubMed:25066128, PubMed:32434865). Localized in phagolysosomes following ingestion of E.coli by neutrophils (PubMed:10947984).
Tissue Location	Bone marrow cells. Neutrophil (PubMed:10947984).