Clone:
REA491
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC, MICS, IF, IHC, ICC, MC
Alternative names:
Myeloperoxidase

Extended validation for MPO Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA491
MPO455-8E6-
CLB-MPO-1-
8E6-
5B8++
Cells were incubated with an excess of purified unconjugated MPO (REA491) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for MPO. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with MPO antibodies. As a control, MPO antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for MPO. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with MPO antibodies. As a control, MPO antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for MPO. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with MPO antibodies. As a control, MPO antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for MPO. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with MPO antibodies. As a control, MPO antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for MPO Antibody, anti-human, REAfinity™

Overview

Clone REA491 recognizes the human myeloperoxidase (MPO) antigen, a heterotetrameric protein consisting of two 15 kDa light chains and two variable-weight glycosylated heavy chains bound to a prosthetic heme group. MPO produces hypochlorous acid from hydrogen peroxide and chloride anion during the neutrophil's respiratory burst. Furthermore, it oxidizes tyrosine to tyrosyl radical using hydrogen peroxide as an oxidizing agent. MPO is present in azurophilic granules of neutrophilic granulocytes and monocytes and is also expressed in some acute myeloid leukemia cells. It plays a major role in the oxygen-dependent microbicidal system of of polymorphonuclear leukocytes. The deficiency of this enzyme activity results in a marked microbicidal defect.
Additional information: Clone REA491 displays negligible binding to Fc receptors.

Alternative names

Myeloperoxidase

Detailed product information

Technical specifications

CloneREA491
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenMPO
Alternative names of antigenMyeloperoxidase
Molecular mass of antigen [kDa]66(sum of molecular weights of subunits)
Distribution of antigengranulocytes, monocytes, neutrophils
Entrez Gene ID4353
RRIDAB_2784435, AB_2752013, AB_2751983, AB_2751873, AB_2751843, AB_2819418, AB_2905337, AB_2905336, AB_2784436

Resources for MPO Antibody, anti-human, REAfinity™

Certificates

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References for MPO Antibody, anti-human, REAfinity™

Publications

  1. Adair, J. E. et al. (2018) Novel lineage depletion preserves autologous blood stem cells for gene therapy of Fanconi anemia complementation group A. Haematologica 103(11): 1806-1814
  2. Manivannan, P. et al. (2014) Can threshold for MPO by flow cytometry be reduced in classifying acute leukaemia? A comparison of flow cytometric and cytochemical myeloperoxidase using different flow cytometric cut-offs. Haematologica : Epub ahead of print
  3. Morishita, K. et al. (1987) Molecular cloning and characterization of cDNA for human myeloperoxidase. J. Biol. Chem. 262(8): 3844-3851
  4. Papayannopoulos, V. et al. (2010) Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps. J. Cell Biol. 191(3): 677-697

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