Mouse MPO ELISA kit

Name: Mouse MPO ELISA kit
SKU: DEIA5691
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Plasma, and cell culture supernatants

Species Reactivity

Mouse

Detection Method

sELISA

Intended Use

The Mouse MPO (Myeloperoxidase) ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse MPO in plasma (serum is not recommended in this assay due to mouse MPO is released from neutrophils into serum in the process of blood coagulation) and cell culture supernatants.

Contents of Kit

1. MPO Microplate (Item A): 96 wells (12 strips × 8 wells) coated with anti-Mouse MPO. Store at 4°C for 1 month. (Return unused wells to the pouch containing desiccant pack, reseal along entire edge.)
2. Wash Buffer Concentrate (20×) (Item B): 25 ml of 20× concentrated solution. Store at 4°C for 1 month.
3. Standard Protein (Item C): 2 vials of Mouse MPO. 1 vial is enough to run each standard in duplicate. Store at -80°C for 1 week.
4. Detection Antibody MPO (Item F): 2 vials of biotinylated anti-Mouse MPO. Each vial is enough to assay half the microplate. Store at 4°C for 5 days.
5. HRP-Streptavidin Concentrate (Item G): 200 μl 400× concentrated HRP-conjugated streptavidin. Do not store and reuse.
6. TMB One-Step Substrate Reagent (Item H): 12 ml of 3,3,5,5'-tetramethylbenzidine (TMB) in buffer solution.
7. Stop Solution (Item I): 8 ml of 0.2 M sulfuric acid.
8. Assay Diluent C (Item L): 30 ml of diluent buffer.
9. Assay Diluent B (Item E): 15 ml of 5× concentrated buffer. Store at 4°C for 1 month.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C. For prepared reagent storage, see table below.

Precision

Intra-Assay CV%: <10%
Inter-Assay CV%: <12%

Detection Range

0.6 ng/ml - 150 ng/ml

Sensitivity

The minimum detectable dose of Mouse MPO was determined to be 600 pg/ml. Minimum detectable dose is defined as the analyte concentration resulting in an absorbance that is 2 standard deviations higher than that of the blank (diluent buffer).

Standard Curve

These standard curves are for demonstration only. A standard curve must be run with each assay.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Microparticles expressing myeloperoxidase as potential biomarkers in anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV)

JOURNAL OF MOLECULAR MEDICINE-JMM

Authors: Manojlovic, M.; Juto, A.; Jonasdottir, A.; Colic, J.; Vojinovic, J.; Nordin, A.; Bruchfeld, A.; Gunnarsson, I; Mobarrez, F.; Antovic, A.

Abstract

To investigate presence of circulating myeloperoxidase-positive microparticles (MPO(+)MPs) in relation to disease activity in patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). Forty-six patients with AAV and 23 age- and sex-matched healthy controls were included. Vasculitis disease activity was assessed using the Birmingham Vasculitis Activity Score (BVAS). MPs were analyzed in citrate plasma by flow cytometry and phenotyped based on MPO expression and co-expression of pentraxin-3 (PTX3), high mobility group box 1 protein (HMGB1), and tumor necrosis factor-like weak inducer of apoptosis (TWEAK). Serum levels of PTX3, sTWEAK, and HMGB1 were also determined. Twenty-three patients had active vasculitis (BVAS >= 1). Concentrations of MPO(+)MPs expressing PTX3, HMGB1, and TWEAK were significantly higher in patients compared to healthy controls (p< 0.001,p< 0.01,p< 0.001, respectively), while concentrations of PTX3(+)and HMGB1(+)MPO(+)MPs were significantly higher in active AAV compared to patients in remission. MPO(+)MPs expressing either PTX3 or HMGB1 were associated with BVAS (r= 0.5,p< 0.001;r= 0.3,p= 0.04, respectively). Significantly higher serum PTX3 levels were found in active- than in inactive AAV (p< 0.001), correlating strongly with BVAS (r= 0.7,p< 0.001). Serum levels of sTWEAK and HMGB1 did not differ between patients and controls. Concentration of MPO(+)MPs is increased in plasma from AAV patients compared to healthy individuals. PTX3 in serum as well as PTX3 and HMGB1 expressed on MPO(+)MPs were associated with disease activity in the investigated patients. Key messages Myeloperoxidase-positive microparticles (MPO+MPs) are increased in plasma from patients with ANCA-associated vasculitis. Concentrations of MPO+MPs expressing PTX3, HMGB1, and TWEAK were significantly higher in patients compared to healthy controls. MPO+MPs expressing PTX3 and HMGB1 are associated with disease activity in ANCA-associated vasculitis.

WKYMVm ameliorates acute lung injury via neutrophil antimicrobial peptide derived STAT1/IRF1 pathway

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

Authors: Lee, Hanbyeol; Lee, Jooyeon; Park, Youngheon; Kim, Jae Ho; Eickelberg, Oliver; Yang, Se-Ran

Abstract

Formyl peptide receptors (FPRs) are mainly expressed on leucocytes and sense microbe-associated molecular pattern (MAMP) molecules, thereby regulating leukocyte chemotaxis and activation. The formyl peptide receptor 2 (FPR2) selective agonist WKYMVm (Trp-Lys-Met-Val-D-Met) has shown potent pro-angiogenic, anti-inflammatory, and anti-apoptotic properties. In this study, we investigated whether WKYMVm exhibits bactericidal activity during neutrophil accumulation in acute lung injury (ALI) in mice and determined its cellular signaling pathways in HL-60 neutrophil-like cells. A daily intraperitoneal treatment of ALI mice with WKYMVm (2.5- and 5 mg/kg/d) daily over four days decreased the levels of proinflammatory cytokines TNF-alpha, IL-6, and IL-1 beta, while it increased the MPO and NO release by differentiated HL-60 neutrophil-like cells. The IRF1 level and STAT1 phosphorylation at 5727 were increased in the lungs of mice with ALI treated with WKYMVm. Lung histology induced by ALI was unaffected by treatment with WKYMVm. In vitro, WKYMVm increased MPO, NO, and SOD activity, as well as IRF1 and STAT1 phosphorylation at Ser727. Taken together, our data suggest therapeutic potential of WKYMVm, via FPR2-dependent regulation of STAT1/IRF1, in ALI. (C) 2020 Elsevier Inc. All rights reserved.