Human Neutrophil Elastase/ELA2 DuoSet ELISA

Product Details
Procedure
Citations (8)
FAQs
Supplemental Products
Reviews (3)

Human Neutrophil Elastase/ELA2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Range
46.9 - 3,000 pg/mL
Sufficient Materials
For five 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Neutrophil Elastase (ELA2). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Human Neutrophil Elastase / ELA2 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Neutrophil Elastase/ELA2

Neutrophil Elastase (ELA2), also known as polymorphonuclear leukocyte elastase, is a serine protease belonging to the chymotrypsin family. Located primarily in the azurophil granules of polymorphonuclear leukocytes, ELA2 function results in the degradation of many extracellular matrix macromolecules. Alpha-1 Antitrypsin (Serpin A1) and secretory leukocyte protease inhibitor (SLPI) have been shown to inhibit ELA2 activity. ELA2 may be involved in lung emphysema, cystic fibrosis, the adult respiratory distress syndrome (ARDS), rheumatoid arthritis, tumor invasion and infectious diseases.

Entrez Gene IDs:
1991 (Human); 50701 (Mouse)
Alternate Names:
Bone marrow serine protease; EC 3.4.21; EC 3.4.21.37; ELA2; ELA2granulocyte-derived elastase; ELANE; elastase 2, neutrophil; elastase, neutrophil expressed; Elastase-2; GE; HLEelastase-2; HNE; Human leukocyte elastase; Leukocyte Elastase; Medullasin; NE; Neutrophil Elastase; PMN elastase; PMN-E; polymorphonuclear elastase; SCN1

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Neutrophil Elastase/ELA2 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. CXCR4 and CXCR7 Inhibition Ameliorates the Formation of Platelet-Neutrophil Complexes and Neutrophil Extracellular Traps through Adora2b Signaling
    Authors: KC Ngamsri, RA Putri, C Jans, K Schindler, A Fuhr, Y Zhang, J Gamper-Tsi, S Ehnert, FM Konrad
    International Journal of Molecular Sciences, 2021;22(24):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Atypical response to bacterial co-infection and persistent neutrophilic broncho-alveolar inflammation distinguish critical COVID-19 from influenza
    Authors: S Cambier, M Metzemaeke, AC Carvalho, A Nooyens, C Jacobs, L Vanderbeke, B Malengier-, M Gouwy, E Heylen, P Meersseman, G Hermans, E Wauters, A Wilmer, C Consortium, D Schols, P Matthys, G Opdenakker, RE Marques, J Wauters, J Vandooren, P Proost
    JCI Insight, 2021;0(0):.
    Species: Human
    Sample Types: BAL Supernatant
  3. Concerted actions by MMPs, ADAMTS and serine proteases during remodeling of the cartilage callus into bone during osseointegration of hip implants
    Authors: J Cassuto, A Folestad, J Göthlin, H Malchau, J Kärrholm
    Bone Rep, 2020;13(0):100715.
    Species: Human
    Sample Types: Plasma
  4. Characterization of ecotin homologs from Campylobacter rectus and Campylobacter showae
    Authors: C Thomas, H Nothaft, R Yadav, C Fodor, A Alemka, O Oni, M Bell, B Rada, CM Szymanski
    PLoS ONE, 2020;15(12):e0244031.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. BCG Vaccination Induces Long-Term Functional Reprogramming of Human Neutrophils
    Authors: SJCFM Moorlag, YA Rodriguez-, J Gillard, S Fanucchi, K Theunissen, B Novakovic, CM de Bont, Y Negishi, ET Fok, L Kalafati, P Verginis, VP Mourits, VACM Koeken, LCJ de Bree, GJM Pruijn, C Fenwick, R van Crevel, LAB Joosten, I Joosten, H Koenen, MM Mhlanga, DA Diavatopou, T Chavakis, MG Netea
    Cell Rep, 2020;33(7):108387.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Role of NOD1/NOD2 receptors in Fusobacterium nucleatum mediated NETosis
    Authors: HM Alyami, LS Finoti, HS Teixeira, A Aljefri, DF Kinane, MR Benakanake
    Microb. Pathog., 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Expressions of MMP-12, TIMP-4, and Neutrophil Elastase in PBMCs and Exhaled Breath Condensate in Patients with COPD and Their Relationships with Disease Severity and Acute Exacerbations
    Authors: W Hao, M Li, Y Zhang, C Zhang, Y Xue
    J Immunol Res, 2019;2019(0):7142438.
    Species: Human
    Sample Types: Serum
  8. G-CSF and GM-CSF Modify Neutrophil Functions at Concentrations found in Cystic Fibrosis
    Authors: S Castellani, S D'Oria, A Diana, AM Polizzi, S Di Gioia, MA Mariggiò, L Guerra, M Favia, A Vinella, G Leonetti, D De Venuto, C Gallo, P Montemurro, M Conese
    Sci Rep, 2019;9(1):12937.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

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Reviews for Human Neutrophil Elastase/ELA2 DuoSet ELISA

Average Rating: 5 (Based on 3 Reviews)

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Human Neutrophil Elastase/ELA2 DuoSet ELISA
By Anonymous on 10/12/2021
Application: Sample Tested: Citrate Plasma

We diluted the plasma 1:100


Human Neutrophil Elastase/ELA2 DuoSet ELISA
By Anonymous on 09/09/2021
Application: Sample Tested: stem cell condition medium

Human Neutrophil Elastase/ELA2 DuoSet ELISA
By Anonymous on 10/16/2019
Application: Sample Tested: EDTA Plasma

We run human EDTA-plasma samples at a 1:50 dilution with high precision.for this analysis.