Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

30

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08246

Entrez-Gene Id:

1991

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:400
Flow Cytometry (Fixed/Permeabilized) 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Neutrophil Elastase (E9C9L) XP® Rabbit mAb recognizes endogenous levels of total neutrophil elastase protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human neutrophil elastase protein.

Background

Neutrophil elastase is a hematopoietic serine protease that belongs to the chymotrypsin superfamily and plays a critical role in the innate immune function of mature neutrophils and monocytes (1,2). Neutrophil elastase is actively synthesized as an inactive zymogen in myelocytic precursor cells of the bone marrow, which then undergoes activation by limited proteolysis and sorting to primary (azurophil) storage granules of mature neutrophil granulocytes for regulated release (3,4). Research studies have shown that neutrophils play a significant role in mediating the inflammatory response through the release of neutrophil elastase, which activates pro-inflammatory cytokines and degrades components of the extracellular matrix and Gram-negative bacteria (5). Mutations in the gene encoding neutrophil elastase, ELA2, have been implicated in hematological diseases such as cyclic and severe congenital neutropenia, which is characterized by defects in promyelocyte maturation (6,7).

  1. Belaaouaj, A. et al. (2000) Science 289, 1185-8.
  2. Tkalcevic, J. et al. (2000) Immunity 12, 201-10.
  3. Gullberg, U. et al. (1999) J Immunol Methods 232, 201-10.
  4. Takahashi, H. et al. (1988) J Biol Chem 263, 2543-7.
  5. Adkison, A.M. et al. (2002) J Clin Invest 109, 363-71.
  6. Horwitz, M. et al. (1999) Nat Genet 23, 433-6.
  7. Bellanné-Chantelot, C. et al. (2004) Blood 103, 4119-25.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#89241

Neutrophil Elastase (E9C9L) XP® Rabbit mAb

Western Blotting Image 1: Neutrophil Elastase (E9C9L) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Neutrophil Elastase (E9C9L) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, neutrophil elastase protein expression is not detected in either A-498 or Jurkat cells.
Western Blotting Image 2: Neutrophil Elastase (E9C9L) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human neutrophil elastase protein (hNE-Myc/DDK; +), using Neutrophil Elastase (E9C9L) XP® Rabbit mAb (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Neutrophil Elastase (E9C9L) XP® Rabbit mAb Expand Image
Immunoprecipitation of neutrophil elastase from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Neutrophil Elastase (E9C9L) XP® Rabbit mAb. Western blot analysis was performed using Neutrophil Elastase (E9C9L) XP® Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used for detection to avoid cross-reactivity with IgG.
Immunofluorescence Image 1: Neutrophil Elastase (E9C9L) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of THP-1 cells differentiated with TPA #4174 (80 nM, 24 hr; left, positive) and A-498 cells (right, negative), using Neutrophil Elastase (E9C9L) XP® Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: Neutrophil Elastase (E9C9L) XP® Rabbit mAb Expand Image
Flow cytometric analysis of Jurkat cells (blue) and U-937 cells (green) using Neutrophil Elastase (E9C9L) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.