Human Myeloperoxidase ELISA Kit - Quantikine

Catalog # Availability Size / Price Qty
DMYE00B
Control Products Available
Best Seller
Human Myeloperoxidase/MPO ELISA Cell Culture Supernate/Cell Lysate/Saliva Standard Curve
3 Images
Product Details
Procedure
Citations (37)
FAQs
Supplemental Products
Reviews (7)

Human Myeloperoxidase Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), Platelet-poor EDTA Plasma (10 uL), Platelet-poor Heparin Plasma (10 uL), Saliva (10 uL), Urine (50 uL)
Sensitivity
0.062 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Urine)
Specificity
Natural human MPO
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Sample Values

Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human MPO in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=29)10021.4-22965
Platelet-poor EDTA plasma (n=29)7.822.89-27.44.93
Platelet-poor heparin plasma (n=29)9.473.57-28.45.27
Saliva (n=10)55831.2-1428461
Urine (n=13)4.390.17-13.54.68

Cell Culture Supernates:
Human monocytes were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 1 μg/mL LPS for 1 and 6 days. Aliquots of the cell culture supernates were removed and assayed for human MPO.

ConditionDay 1 (ng/mL)Day 6 (ng/mL)
Unstimulated28.041.4
Stimulated29.930.3

HL-60 human acute promyelocytic leukemia cells were cultured in RPMI supplemented with 20% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human MPO, and measured 18.2 ng/mL.

An aliquot of the same HL-60 cells was removed, and the cells were lysed at 5 x 106 cells/mL in diluted Cell Lysis Buffer 3. The lysate was assayed for human MPO and measured 2673 ng/mL.


Product Summary

The Quantikine Human MPO Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human MPO in cell culture supernates, cell lysates, serum, platelet-poor plasma, saliva, and urine.

Recovery

The recovery of MPO spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 92-105
Platelet-poor EDTA Plasma (n=4) 95 90-101
Platelet-poor Heparin Plasma (n=4) 91 83-100
Serum (n=4) 100 88-110

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of MPO were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Myeloperoxidase/MPO ELISA Linearity

Scientific Data

Human Myeloperoxidase/MPO ELISA Cell Culture Supernate/Cell Lysate/Saliva Standard Curve

Human Myeloperoxidase/MPO ELISA Serum/Plasma/Urine Standard Curve

Product Datasheets

You must select a language.

x

Preparation and Storage

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

Background: Myeloperoxidase/MPO

Myeloperoxidase (MPO) is a heme-containing enzyme belonging to the XPO subfamily of peroxidases. It is an abundant neutrophil and monocyte glycoprotein that catalyzes the hydrogen peroxide dependent formation of hypochlorus acid (HOCl) and other reactive species. Enzymatically active MPO is a disulfide-linked tetramer that contains two heme groups and two copies each of the heavy and light chains. MPO binds Albumin, MMR, Cytokeratin 1 on vascular endothelial cells, HMW Kininogen, and Integrin CD11b/CD18 on neutrophils. These interactions promote MPO clearance, a reduction of nitric oxide and bradykinin levels, reduced vasodilation, and continued neutrophil activation. Neutrophil MPO is stored in cytoplasmic azurophilic granules. Upon cellular activation and degranulation, MPO is delivered into phagosomes where it is required for the killing of phagocytosed bacteria. Activated neutrophils also release granule contents extracellularly. Elevated plasma MPO levels have been associated with a variety of clinical conditions including systemic inflammation, eclampsia, risk of cardiovascular events, vascular endothelial dysfunction, severity of multiple sclerosis, and prospective mortality and oxidative stress during hemodialysis.

Entrez Gene IDs:
4353 (Human); 17523 (Mouse); 303413 (Rat)
Alternate Names:
EC 1.11.1; EC 1.11.1.7; MPO; Myeloperoxidase
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. Add Assay Diluent
  4.   For Serum, Plasma, & Human Milk Samples: Add 50 µL of Assay Diluent to each well.
    For Cell Culture Supernate, Cell Lysate, Saliva, & Urine Samples: Add 100 µL Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well.
  10.   For Serum, Plasma, & Human Milk Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    For Cell Culture Supernate, Cell Lysate, Saliva, & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
  11.   Aspirate and wash 4 times.

  12. 200 µL Substrate Solution
  13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes on the benchtop. PROTECT FROM LIGHT.

  14. 50 µL Stop Solution
  15. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Myeloperoxidase Quantikine ELISA Kit

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.

37 Citations: Showing 1 - 10
Filter your results:

Filter by:

  1. Prognostic value of coronary atherosclerosis and CAC score for the risk of chemotherapy-related cardiac dysfunction (CTRCD): The protocol of ANTEC study
    Authors: Borowiec, A;Ozdowska, P;Rosinska, M;Jagiello-Gruszfeld, A;Jasek, S;Waniewska, J;Kotowicz, B;Kosela-Paterczyk, H;Lampka, E;Makowka, A;Fuksiewicz, M;Chojnacka, M;Zebrowska, A;Gepner, K;Kapala, A;Cieszanowski, A;Nowecki, Z;Walewski, J;
    PloS one
    Species: Human
    Sample Types: Plasma
  2. Serum Levels of Proinflammatory Biomarkers in Military Recruits with and without Metabolic Syndrome
    Authors: Al Asmari, AK;Al Shehri, HA;Khan, HA;Al Omani, S;Kadasah, SG;Horaib, GB;Al Buraidi, A;Al Sharif, AA;Mohammed, FS;Abbasmanthiri, R;Osman, NM;
    Mediators of inflammation
    Species: Human
    Sample Types: Serum
  3. Blood concentrations of mediators released from activated neutrophils are related to the severity of alcohol-induced liver damage
    Authors: B Kasztelan-, B Zygo, A Rycyk-Boja, A Surdacka, J Rolinski, H Cichoz-Lac
    PLoS ONE, 2023-01-06;18(1):e0280068.
    Species: Human
    Sample Types: Serum
  4. NETosis and Nucleosome Biomarkers in Septic Shock and Critical COVID-19 Patients: An Observational Study
    Authors: L Morimont, M Dechamps, C David, C Bouvy, C Gillot, H Haguet, J Favresse, L Ronvaux, J Candiracci, M Herzog, PF Laterre, J De Poorter, S Horman, C Beauloye, J Douxfils
    Biomolecules, 2022-07-27;12(8):.
    Species: Human
    Sample Types: Plasma
  5. Rare germline alterations of myeloperoxidase predispose to myeloid neoplasms
    Authors: S Kongkiatka, L Terkawi, Y Guan, V Adema, M Hasipek, T Dombrovski, M Co, W Walter, H Awada, Y Parker, S Hutter, S Pagliuca, C Gurnari, HJ Rogers, M Meggendorf, DJ Lindner, T Haferlach, V Visconte, T LaFrambois, BK Jha, JP Maciejewsk
    Leukemia, 2022-06-27;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  6. Cell-Free DNA, Neutrophil extracellular traps (NETs), and Endothelial Injury in Coronavirus Disease 2019- (COVID-19-) Associated Acute Kidney Injury
    Authors: BM Henry, MHS de Oliveir, I Cheruiyot, J Benoit, J Rose, EJ Favaloro, G Lippi, S Benoit, N Pode Shakk
    Mediators of Inflammation, 2022-04-22;2022(0):9339411.
    Species: Human
    Sample Types: Plasma
  7. Tumour cell apoptosis modulates the colorectal cancer immune microenvironment via interleukin-8-dependent neutrophil recruitment
    Authors: V Schimek, K Strasser, A Beer, S Göber, N Walterskir, C Brostjan, C Müller, T Bachleitne, M Bergmann, H Dolznig, R Oehler
    Cell Death & Disease, 2022-02-04;13(2):113.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Association of Lipoproteins with Neutrophil Extracellular Traps in Patients with Abdominal Aortic Aneurysm
    Authors: A Brandau, N Ibrahim, J Klopf, H Hayden, M Ozsvar-Koz, T Afonyushki, S Bleichert, L Fuchs, V Watzinger, V Nairz, E Manville, V Kessler, H Stangl, W Eilenberg, C Neumayer, C Brostjan
    Biomedicines, 2022-01-20;10(2):.
    Species: Human
    Sample Types: Plasma
  9. Association of circulating MR-proADM with all-cause and cardiovascular mortality in the general population: Results from the KORA F4 cohort study
    Authors: C Gar, B Thorand, C Herder, C Sujana, M Heier, C Meisinger, A Peters, W Koenig, W Rathmann, M Roden, M Stumvoll, H Maalmi, T Meitinger, H Then, J Seissler, C Then
    PLoS ONE, 2022-01-06;17(1):e0262330.
    Species: Human
    Sample Types: Serum
  10. NETs biomarkers in saliva and serum OSCC patients: One hypothesis, two conclusions
    Authors: M Garley, D Dziemia?cz, W Ratajczak-, A Pryczynicz, K Nowak, B ?azarczyk, E Jab?o?ska
    Advances in medical sciences, 2021-12-28;67(1):45-54.
    Species: Human
    Sample Types: Saliva
  11. Role of Clinical Characteristics and Biomarkers at Admission to Predict One-Year Mortality in Elderly Patients with Pneumonia
    Authors: A Malézieux-, L Azurmendi, S Pagano, N Vuilleumie, JC Sanchez, D Zekry, JL Reny, J Stirnemann, N Garin, V Prendki, On Behalf
    Journal of Clinical Medicine, 2021-12-25;11(1):.
    Species: Human
    Sample Types: Plasma
  12. Association between Biomarkers of Oxidative Stress and Inflammation with Cardiac Necrosis and Heart Failure in Non-ST Segment Elevation Myocardial Infarction Patients and Various Degrees of Kidney Function
    Authors: S Roumelioti, A Veljkovic, PI Georgianos, G Lazarevic, Z Perisic, J Hadzi-Djok, V Liakopoulo, G Kocic
    Oxidative Medicine and Cellular Longevity, 2021-11-01;2021(0):3090120.
    Species: Human
    Sample Types: Serum
  13. ELISA detection of MPO-DNA complexes in human plasma is error-prone and yields limited information on neutrophil extracellular traps formed in vivo
    Authors: H Hayden, N Ibrahim, J Klopf, B Zagrapan, LM Mauracher, L Hell, TM Hofbauer, AS Ondracek, C Schoergenh, B Jilma, IM Lang, I Pabinger, W Eilenberg, C Neumayer, C Brostjan
    PLoS ONE, 2021-04-22;16(4):e0250265.
    Species: Human
    Sample Types: Plasma
  14. Profile of new vascular damage biomarkers in middle-aged men with arterial hypertension
    Authors: AE Charkiewic, M Garley, W Ratajczak-, K Nowak, E Jab?o?ska, D Ma?lach, WJ Omeljaniuk
    Advances in medical sciences, 2021-03-05;66(1):185-191.
    Species: Human
    Sample Types: Serum
  15. Characterization of ADAMTS13 and von Willebrand factor levels in septic and non-septic ICU patients
    Authors: K Singh, AC Kwong, H Madarati, S Kunasekara, T Sparring, AE Fox-Robich, PC Liaw, CA Kretz
    PLoS ONE, 2021-02-19;16(2):e0247017.
    Species: Human
    Sample Types: Plasma
  16. Granulocyte and monocyte adsorptive apheresis induces apoptosis of neutrophils and release of a novel chemoattractant for desensitization of interleukin-8 response - In vitro and in vivo results
    Authors: N Kashiwagi, F Saito, H Maegawa, K Kaneda
    Cytokine, 2021-01-01;139(0):155410.
    Species: Human
    Sample Types: Plasma
  17. Resveratrol and exercise combined to treat functional limitations in late life: A pilot randomized controlled trial
    Authors: SA Harper, JR Bassler, S Peramsetty, Y Yang, LM Roberts, D Drummer, RT Mankowski, C Leeuwenbur, K Ricart, RP Patel, MM Bamman, SD Anton, BC Jaeger, TW Buford
    Exp Gerontol, 2020-10-15;0(0):111111.
    Species: Human
    Sample Types: Plasma
  18. The role of specific biomarkers, as predictors of post-operative complications following flexible ureterorenoscopy (FURS), for the treatment of kidney stones: a single-centre observational clinical pilot-study in 37 patients
    Authors: SF Hughes, AJ Moyes, RM Lamb, P Ella-Tongw, C Bell, A Moussa, I Shergill
    BMC Urol, 2020-08-14;20(1):122.
    Species: Human
    Sample Types: Plasma, Serum
  19. Biomarkers of neutrophil extracellular traps (NETs) and nitric oxide-(NO)-dependent oxidative stress in women who miscarried
    Authors: WJ Omeljaniuk, E Jab?o?ska, M Garley, A Pryczynicz, W Ratajczak-, K Socha, MH Borawska, AE Charkiewic
    Sci Rep, 2020-08-04;10(1):13088.
    Species: Human
    Sample Types: Serum
  20. Neutrophil Phenotypes in Coronary Artery Disease
    Authors: P Maréchal, J Tridetti, ML Nguyen, O Wéra, Z Jiang, M Gustin, AF Donneau, C Oury, P Lancellott
    J Clin Med, 2020-05-25;9(5):.
    Species: Human
    Sample Types: Plasma
  21. Neutrophil activation and NETosis are the major drivers of thrombosis in heparin-induced thrombocytopenia
    Authors: J Perdomo, HHL Leung, Z Ahmadi, F Yan, JJH Chong, FH Passam, BH Chong
    Nat Commun, 2019-03-21;10(1):1322.
    Species: Human
    Sample Types: Plasma
  22. Neutrophil Dysfunction in the Airways of Children with Acute Respiratory Failure Due to Lower Respiratory Tract Viral and Bacterial Coinfections
    Authors: JR Grunwell, VD Giacalone, S Stephenson, C Margaroli, BS Dobosh, MR Brown, AM Fitzpatric, R Tirouvanzi
    Sci Rep, 2019-02-27;9(1):2874.
    Species: Human
    Sample Types: Tracheal Aspirate
  23. Cardiac magnetic resonance using late gadolinium enhancement and atrial T1 mapping predicts poor outcome in patients with atrial fibrillation after catheter ablation therapy
    Authors: JA Luetkens, AC Wolpers, T Beiert, D Kuetting, D Dabir, R Homsi, H Meenderman, NA Dayé, V Knappe, M Karsdal, SH Nielsen, F Genovese, F Stöckigt, M Linhart, D Thomas, G Nickenig, HH Schild, JW Schrickel, RP Andrié
    Sci Rep, 2018-09-11;8(1):13618.
    Species: Human
    Sample Types: Serum
  24. Myeloperoxidase level and inflammatory markers and lipid and lipoprotein parameters in stable coronary artery disease
    Authors: E Kimak, B Zi?ba, D Duma, J Solski
    Lipids Health Dis, 2018-04-04;17(1):71.
    Species: Human
    Sample Types: Serum
  25. Citrullinated histone H3 as a novel prognostic blood marker in patients with advanced cancer
    Authors: C Thålin, S Lundström, C Seignez, M Daleskog, A Lundström, P Henriksson, T Helleday, M Phillipson, H Wallén, M Demers
    PLoS ONE, 2018-01-11;13(1):e0191231.
    Species: Human
    Sample Types: Plasma
  26. Paraoxonase-1 and myeloperoxidase correlate with vascular biomarkers in overweight patients with newly diagnosed untreated hyperlipidaemia
    Authors: A Szentpéter, N Zsíros, VE Varga, H L?rincz, M Katkó, I Seres, P Fülöp, G Paragh, M Harangi
    VASA, 2017-06-12;0(0):1-7.
    Species: Human
    Sample Types: Serum
  27. Inflammatory Differences in Plaque Erosion and Rupture in Patients With ST-Segment Elevation Myocardial Infarction
    Authors: S Chandran, J Watkins, A Abdul-Aziz, M Shafat, PA Calvert, KM Bowles, MD Flather, SA Rushworth, AD Ryding
    J Am Heart Assoc, 2017-05-03;6(5):.
    Species: Human
    Sample Types: Plasma
  28. The interdependencies of viral load, the innate immune response, and clinical outcome in children presenting to the emergency department with respiratory syncytial virus-associated bronchiolitis
    Authors: FA Piedra, M Mei, V Avadhanula, R Mehta, L Aideyan, RP Garofalo, PA Piedra
    PLoS ONE, 2017-03-07;12(3):e0172953.
    Species: Human
    Sample Types: Nasal Lavage Fluid
  29. Complex regulation of neutrophil-derived MMP-9 secretion in central nervous system tuberculosis
    Authors: CW Ong, PJ Pabisiak, S Brilha, P Singh, F Roncaroli, PT Elkington, JS Friedland
    J Neuroinflammation, 2017-02-07;14(1):31.
    Species: Human
    Sample Types: Whole Cells
  30. Combined administration of anisodamine and neostigmine rescued acute lethal crush syndrome through ?7nAChR-dependent JAK2-STAT3 signaling
    Sci Rep, 2016-11-22;6(0):37709.
    Species: Mouse
    Sample Types: Serum
  31. Inflammatory thresholds and the species-specific effects of colonising bacteria in stable chronic obstructive pulmonary disease.
    Authors: Singh R, Mackay A, Patel A, Garcha D, Kowlessar B, Brill S, Donnelly L, Barnes P, Donaldson G, Wedzicha J
    Respir Res, 2014-09-14;15(0):114.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. Growth differentiation factor-15 (GDF-15) levels are associated with cardiac and renal injury in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass.
    Authors: Kahli A, Guenancia C, Zeller M, Grosjean S, Stamboul K, Rochette L, Girard C, Vergely C
    PLoS ONE, 2014-08-29;9(8):e105759.
    Species: Human
    Sample Types: Plasma
  33. Increased levels of leukocyte-derived MMP-9 in patients with stable angina pectoris.
    Authors: Jonasson S, Lundberg A, Kalvegren H, Bergstrom I, Szymanowski A, Jonasson L
    PLoS ONE, 2011-04-29;6(4):e19340.
    Species: Human
    Sample Types: Serum
  34. Presence of IL-5 protein and IgE antibodies to staphylococcal enterotoxins in nasal polyps is associated with comorbid asthma.
    Authors: Bachert C, Zhang N, Holtappels G, De Lobel L, Van Cauwenberge P, Liu S, Lin P, Bousquet J, Van Steen K
    J. Allergy Clin. Immunol., 2010-11-01;126(5):962.
    Species: Human
    Sample Types: Tissue Homogenates
  35. Interleukin-8 is increased in the membrane of circulating erythrocytes in patients with acute coronary syndrome.
    Authors: Tziakas DN, Chalikias GK, Tentes IK, Stakos D, Chatzikyriakou SV, Mitrousi K, Kortsaris AX, Kaski JC, Boudoulas H
    Eur. Heart J., 2008-09-02;29(22):2713-22.
    Species: Human
    Sample Types: Plasma
  36. Effects of atorvastatin added to inhaled corticosteroids on lung function and sputum cell counts in atopic asthma.
    Authors: Hothersall EJ, Chaudhuri R, McSharry C, Donnelly I, Lafferty J, McMahon AD, Weir CJ, Meiklejohn J, Sattar N, McInnes I, Wood S, Thomson NC
    Thorax, 2008-08-29;63(12):1070-5.
    Species: Human
    Sample Types: Sputum
  37. Computed tomographic scan-diagnosed chronic obstructive pulmonary disease-emphysema: eotaxin-1 is associated with bronchodilator response and extent of emphysema.
    Authors: Miller M, Ramsdell J, Friedman PJ, Cho JY, Renvall M, Broide DH
    J. Allergy Clin. Immunol., 2007-11-01;120(5):1118-25.
    Species: Human
    Sample Types: BALF

FAQs

  1. Does the Human Myeloperoxidase Quantikine ELISA Kit work with sputum samples?

    • We have not tested sputum samples in this kit. We are aware that customers have cited the use of this kit with sputum samples. See the Citations tab. Additional sample validation by the investigator may be required.

View all ELISA FAQs
Loading...

Reviews for Human Myeloperoxidase Quantikine ELISA Kit

Average Rating: 4.6 (Based on 7 Reviews)

5 Star
57.14%
4 Star
42.86%
3 Star
0%
2 Star
0%
1 Star
0%

Have you used Human Myeloperoxidase Quantikine ELISA Kit?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review

Filter by:


Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 10/16/2023
Sample Tested: Plasma

Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 11/14/2022
Sample Tested: Urine

Urine sample should maybe be concentrated


Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 11/06/2022
Sample Tested: Serum

Human Myeloperoxidase Quantikine ELISA Kit
By Melania Guerrero-Hue on 11/02/2022
Sample Tested: Serum

Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 06/09/2022
Sample Tested: Serum

Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 11/05/2018
Sample Tested: Cell culture supernatant

Human Myeloperoxidase Quantikine ELISA Kit
By Anonymous on 11/02/2018
Sample Tested: Cell culture supernatant,Neutrophils