Anti-Neutrophil Elastase/ELANE Antibody

• Catalog Number: orb381060
• Category: Antibodies
• Description: Anti-Neutrophil Elastase/ELANE Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived mouse Elastase/ELANE/ELA2 recombinant protein (Position: S27-N265). Mouse Elastase/ELANE/ELA2 shares 92% amino acid (aa) sequence identity with rat Elastase/ELANE/ELA2.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 29 kDa
• UniProt ID: Q3UP87
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Neutrophil elastase; 3.4.21.37; Elastase-2; Leukoc
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Mouse Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat ELISA, 0.1-0.5μg/ml,-. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody. Elastase/ELANE/ELA2 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Elastase/ELANE/ELA2 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody. Elastase/ELANE/ELA2 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Elastase/ELANE/ELA2 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody. 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: mouse bone tissue lysates, Lane 2: mouse spleen tissue 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-Elastase/ELANE/ELA2 antigen affinity purified polyclonal antibody at 0.5 µg/mL 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Elastase/ELANE/ELA2 at approximately 29 kDa. The expected band size for Elastase/ELANE/ELA2 is at 29 kDa.