Invitrogen

Neutrophil elastase Recombinant Rabbit Monoclonal Antibody (JF098-6)

View all (27) Neutrophil elastase antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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Neutrophil elastase Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Neutrophil elastase was performed using THP-1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Neutrophil elastase Recombinant Rabbit Monoclonal Antibody (JF098-6) (Product # MA5-32548) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Neutrophil elastase Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of Neutrophil elastase of paraffin-embedded Human spleen tissue using a Neutrophil-elastase Monoclonal antibody (Product #MA5-32548). Counter stained with hematoxylin.

Neutrophil elastase Antibody in Western Blot (WB) — Western blot was performed using Anti-Neutrophil elastase Recombinant Rabbit Monoclonal Antibody (JF098-6) (Product # MA5-32548) and a 29kDa band corresponding to Neutrophil elastase was observed in HL-60, U-937, THP-1 but not in HEk-293, MDA-MB-231 and A549 which are reported to be negative. Whole cell extracts of HL-60 (Lane 1), U-937 (Lane 2), THP-1 (Lane 3), HEK-293 (Lane 4), MDA-MB-231 (Lane 5) and A549 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). An uncharacterized band was also observed at ~15kDa.

Neutrophil elastase Antibody in Flow Cytometry (Flow) — Flow Cytometric analysis of Neutrophil elastase in HepG2 cells using a Neutrophil elastase Monoclonal Antibody (Product # MA5-32548) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.

Product Details

MA5-32548

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

Immunohistochemistry (IHC)

1:50-1:200

Immunocytochemistry (ICC/IF)

1:50

Flow Cytometry (Flow)

1:10-1:50

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Expression System

HEK293 cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

JF098-6

Immunogen

Recombinant protein within Human Neutrophil Elastase aa 1-140

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 40% Glycerol, 0.05% BSA

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2809825

Product Specific Information

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

The neutrophil form of elastase is 218 amino acids long, with two asparagine-linked carbohydrate chains (see glycosylation). It is present in azurophil granules in the neutrophil cytoplasm. There appear to be two forms of neutrophil elastase, termed IIa and IIb.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Bone marrow serine protease; ELA2A; Elastase; elastase 2, neutrophil; Elastase-2; Elastase2; granulocyte-derived elastase; HLE; Human leukocyte elastase; leukocyte elastase; Medullasin; Neutrophil elastase; PMN elastase; polymorphonuclear elastase

Gene Aliases: ELA2; ELANE; GE; HLE; HNE; NE; PMN-E; SCN1

UniProt ID: (Human) P08246

Entrez Gene ID: (Human) 1991

Molecular Function: serine protease

Function(s)

negative regulation of transcription from RNA polymerase II promoter response to yeast acute inflammatory response to antigenic stimulus proteolysis cellular calcium ion homeostasis phagocytosis response to UV extracellular matrix disassembly protein catabolic process response to lipopolysaccharide defense response to bacterium positive regulation of MAP kinase activity negative regulation of growth of symbiont in host negative regulation of chemokine biosynthetic process negative regulation of interleukin-8 biosynthetic process positive regulation of interleukin-8 biosynthetic process positive regulation of smooth muscle cell proliferation negative regulation of inflammatory response positive regulation of immune response leukocyte migration negative regulation of chemotaxis neutrophil mediated killing of fungus

Process(es)

RNA polymerase II transcription corepressor activity protease binding endopeptidase activity serine-type endopeptidase activity protein binding heparin binding peptidase activity cytokine binding