PA5-16672

antibody from Invitrogen Antibodies

Targeting: MPO

Provider product page for PA5-16672

ELISA

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [89]
Comments [0]
Validations
Immunohistochemistry [3]
Other assay [23]

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Product number: PA5-16672 - Provider product page
Provider: Invitrogen Antibodies
Product name: Myeloperoxidase Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Purifed from natural sources
Description: PA5-16672 targets Myeloperoxidase in IHC (P) applications and shows reactivity with Human, mouse, and Rat samples. The PA5-16672 immunogen is purified human granulocytic MPO. PA5-16672 detects MPO-DNA complexes in purified PMA-induced human neutrophil extracellular traps in ELISA application. Concentration is not determined for this particular antibody. The final product is manufactured from stock material based upon a dilution ratio. The antibody is still quality tested to ensure expected performance and lot-to-lot consistency.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 500 µL
Storage: 4° C

MPO protein structure - PA5-16672 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: A 96-well ELISA plate was coated with capture antibody (anti-MPO, Product # PA5-16672, diluted 1:000) in PBS overnight. After blocking with 1% BSA, purified PMA-induced human neutrophil extracellµlar traps (NETs, induced as described in Lood C et al Nat Med 2016) were added in 1% BSA in PBS with 2 mM EDTA overnight. As a detection antibody, anti-DNA-POD diluted 1:100 was used, and the ELISA was developed with TMB substrate. Resµlts are presented as absorbance units at 450 nm. As indicated in the figure, NETs (containing MPO-DNA complexes) were readily detected by the MPO antibody, whereas the controls (either adding PBS instead of NETs, or not adding the detection antibody) gave a low absorbance reading. Data courtesy of Thermo Fisher Antibody Data Exchange Program.

Supportive validation

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Experimental details: Figure 3 Ferulic acid reduced the level of advanced oxidation protein products (AOPPs) and myeloperoxidase (MPO) in MN mouse model. At the end of the experiment, serum and renal cortical tissues were collected, and renal cortical tissues were processed by sonicator in a ratio of per mg tissue in 5 mu L phosphate buffer. The formation of oxidation protein was determined by detecting the AOPPs levels in serum (a) and renal cortical tissues (b). Paraffin-embedded mouse kidney sections were examined their expression of MPO by immunohistochemical staining in NC ((c) and (g)), N-T ((d) and (h)), Pre-T ((e) and (i)) and Post-T ((f) and (j)) groups. Original magnifications: 400x ((c), (d), (e), and (f)) and 40x ((g), (h), (i), and (j)) (* P < 0.05; ** P < 0.01).

Supportive validation

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Experimental details: Figure 4 Myeloperoxidase positive cells are present in much greater numbers in GC-C -/- IL-10 -/- colonic tissue as compared to IL-10 -/- . Representative images showing greater numbers of MPO stained cells, most of which are neutrophils, in colon lacking GC-C and IL-10. Myeloperoxidase is depicted in green while nuclei are stained blue using DAPI. Magnification = 200X.

Supportive validation

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Experimental details: Fig. 5 Immunohistochemistry for macrophages: F4/80 (A and B), for neutrophils: myeoperoxidase (D and E) and for T lymphocytes: CD3 (G and H) in livers of control (A, D, G) and caAlk5/Alb-Cre mice (B, E, H) treated with TAA for 12 weeks. Quantification of F4/80-(C), myeloperoxidase-(F), and CD3-(I) positive cells in livers from saline-treated and TAA-treated mice. Data represent means +- SD, n = 5 mice per experimental group, ten 20x fields per mouse. There is a significant difference after TAA treatment when compared with genotype-specific saline control (*P < 0.05). There is a significant difference when compared with genotype-specific saline control and between genotypes after TAA treatment (**P < 0.05). Immunochemistry with NovaRed substrate, counterstained with hematoxylin. CV denotes the central vein. PT denotes the portal tract. Fig. 5

Supportive validation

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Experimental details: Figure 1 Acute allograft transplant rejection induces graft neutrophil PE and the PGP protease cascade. Eight- to ten-week-old, male, BALB/c mice received heterotopic heart transplants from similarly aged BALB/c (isograft) or C57BL/6 (allograft) donor mice. Mice were sacrificed 3 days after transplant. Among allograft mice, laser confocal microscopy revealed colocalization of PE with the neutrophil marker myeloperoxidase within the extracellular matrix ( A ). This neutrophilic response was further quantified in Supplemental Figure 1 . Importantly, both total expression of the PGP-generating metalloprotease PE ( B ) ( n = 5) and PE activity ( C ) ( n = 7 and n = 5, respectively) were increased within allograft hearts and associated with de novo PGP production ( D ) ( n = 5). Finally, increased myocardial PGP production was positively associated with ET-1 ( E ) ( n = 6), an established marker of PGP activity. Results presented as mean +- SE. Where indicated, n represents animals/group. Student's unpaired t test was used for each comparison. PE, prolyl endopeptidase; PGP, proline-glycine-proline; ET-1, endothelin-1.

Supportive validation

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Experimental details: Figure 2 PE inhibition attenuates allograft inflammatory response. Allograft mice were treated with ZPP (10 mg/kg i.p.) or a 2% DMSO vehicle control. ( A-F ) illustrates representative images of H&E and IHC staining. Compared with controls ( A-C ), ZPP reduced allograft inflammatory cell invasion ( D and E ) and MPO expression ( F ). Although total allograft PE did not change ( G ) ( n = 5 animals/group), ZPP administration significantly reduced serum PE activity ( H ) ( n = 5 DMSO and n = 6 ZPP). Importantly, ZPP significantly reduced allograft PGP ( I ) ( n = 5 DMSO and n = 6 ZPP) and ET-1 ( J ) ( n = 5 animals/group). This observation was associated with reduced intragraft neutrophil invasion ( K ) ( n = 5 animals/group). In contrast, concentrations of the antifibrotic peptide AcSDKP were not altered by the administration of ZPP ( L ) ( n = 5 DMSO and n = 6 ZPP animals/group). Importantly, ZPP attenuated the expression of allograft IFN-gamma and associated leukocyte chemoattractants CXCL10 and RANTES ( M-O ) ( n = 5 DMSO and n = 7 ZPP). Results presented as mean +- SE. Student''s unpaired t test was used for each comparison. PE, prolyl endopeptidase; PGP, proline-glycine-proline; ET-1, endothelin-1; ZPP, Z-Pro-Prolinal; MPO, myeloperoxidase; AcSDKP, acetyl-N-Ser-Asp-Lys-Pro.

Supportive validation

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Experimental details: Figure 4 Effect of clodronate liposomes on Prx2-induced neutrophil infiltration. ( a ) Examples of myeloperoxidase (MPO) immunohistochemistry at the choroid plexus one day after icv Prx2 + clodronate liposome (Prx2 + clo) and Prx2 + control liposome (Prx2 + control) injections. ( b ) The numbers of MPO positive cells were quantified as a % of all choroid plexus cells. Values are the mean +- SD; n = 7; # p < 0.01 vs. control group. Scale bar = 50 mum at low magnification, 20 mum at high magnification. Dots represent data for each animal.

Supportive validation

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Experimental details: Figure 1. Immune cell frequencies a and b immunohistochemistry (IHC) based identification of myeloperoxidase (MPO) and CD14 expressing cells, respectively. A section from each tumor was stained for either MPO or CD14, images at 10x were taken from 5 different regions on each slide and the ratio of MPO/CD14 stain to the DAPI (nuclear stain) was determined using image analysis. Ratios (individual samples, and combined based on tumor grading) are shown in box plots. Images of three sections representing a low, medium and high ratio are shown as an inset in the graphs. Top panel of insets are images of IHC, and the bottom panel are digital conversions with pink representing the marker and blue nuclei for ease of viewing (enhanced contrast). In addition, percentages of CD15 + (neutrophils) and CD14 + (monocytes) cells determined by flow cytometry are shown below for comparison. c percentages of immune cell subsets in tumors determined via flow cytometry. Percentages are calculated as the proportion of each subset among total live cells. Cell types were determined using the following markers: CD45 + - all immune cells; CD15 high - neutrophils; CD15 med - other granulocytes (gran. ); and CD14 + CD15 neg and CD14 + CD15 med as two monocyte subsets. d percentages of immune cell subsets in the blood. CD15 high - neutrophils and CD14 + CD15 neg - monocytes. For statistical comparison of data, two-way ANOVA followed by Bonferroni's test was performed. *** indicates p < .001. No significa

Supportive validation

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Experimental details: Figure 4 Sesn1/2/3 TKO mice manifest hepatic inflammation on WD. ( A ) Immunofluorescence analysis of MPO and F4/80 in liver sections of WD-treated mice (original magnification, x100). ( B ) Quantification of MPO- and F4/80-positive cells in ( A ). ( C and D ) Real-time PCR analysis of inflammation-related genes in livers of control diet or WD treated mice. ( E-H ) Hepatic cytokine levels were measured using enzyme-linked immunosorbent assay kits. Data are expressed as mean +- SD (n = 4-5). * P < .05, ** P < .01, and *** P < .001 for the respective genotype on WD vs control diet.

Supportive validation

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Experimental details: Figure 5 Absence of the Pad2 gene inhibits NET formation both in vivo and in vitro. WT and Pad2 -/- mice were subjected to CLP. ( A and B ) The concentrations of dsDNA in mouse serum ( n = 3 for WT mice without CLP group, n = 5 for WT mice with CLP group, n = 3 for Pad2 -/- mice without CLP group, n = 7 for Pad2 -/- mice with CLP group) ( A ) and peritoneal lavage ( n = 4 for WT mice without CLP group, n = 9 for WT mice with CLP group, n = 3 for Pad2 -/- mice without CLP group, and n = 8 for Pad2 -/- mice with CLP group) ( B ) after 24 hours. BM neutrophils were isolated from WT and Pad2 -/- mice and were treated with A23187 or PMA. ( C ) NET formation in response to calcium ionophore and PMA in WT and Pad2 -/- neutrophils at different time points ( n = 3-5/group). ( D ) Representative immunofluorescence images of the NET formation in WT and Pad2 -/- neutrophils after vehicle, Ca 2+ ionophore (A23187), or PMA treatment for 3.5 hours. ( E and F ) Quantification results of immunofluorescence images ( n = 6/group). ( G ) Levels of myeloperoxidase-DNA (MPO-DNA) in cell supernatant of WT and Pad2 -/- neutrophils after vehicle, A23187or PMA treatment for 3.5 hours. Results are representative of 2 independent experiments ( C - G ). Data are expressed as mean +- SEM. Data were analyzed by 2-way ANOVA with Bonferroni's multiple comparisons test ( A - C and E - G ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bar: 50 mum. Ve, vehicle.

Supportive validation

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Experimental details: 10.1371/journal.pone.0242318.g002 Fig 2 Effects of colchicine therapy on lung injury, neutrophils recruitment, and NETosis after oleic acid-induced ALI. Values are mean +- SEM. **** p

Supportive validation

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Experimental details: Fig. 1 iPC injection mitigates immune response as compared to IM injection in rats. a Schematic illustration of iPC injection and in situ formation of cardiac patch. b H&E staining showing the formation of a cardiac patch 7 days after iPC injection in the iPC-iPS-CPC group (dash lines delineate the injected cells). Intramyocardial (IM) injection of iPS-CPCs in ECM hydrogel caused massive infiltration of immune cells (arrows). iPC injection reduced immune cell infiltration and promoted regional formation of blood vessels (asterisks) in the patch. c Representative microscopic images showing the presence of neutrophils, CD4 and CD8 T cells, and corresponding quantitation. Scale bar, 60 mum. Data were shown as mean +- SD, n = 4 animals per group. Comparison of two groups was performed with the unpaired, two-tailed Student's t -test, ** p < 0.01. Source data are provided in a Source Data file. MI myocardial infarction, iPC intrapericardial injection, IM intramyocardium injection, iPS-CPC induced pluripotent stem cell-derived cardiac progenitor cell, ECM extracellular matrix, MPO myeloperoxidase, CD cluster of differentiation, HPF high-power field.

Supportive validation

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Experimental details: Figure 3. Chronic gastritis in GEC C1galt1 -/- mice is driven by mucosal inflammasomes. (A ) Western blot analysis of stomach lysates. p10 is the activated form of Casp1. (B) Densitometry of the Casp1 p10 subunit visualized via Western blot of stomach lysates over time. (C) Enzyme-linked immunosorbent assay of gastric organ culture supernatants (mean +- SD, n = 5 mice/group). (D) IF staining for Casp1 in gastric antrum sections. Arrow indicates carcinoma. Scale bars, 50 um. (E) Generation of inflammasome-deficient GEC C1galt1 -/- mice. (F) Representative macroscopic and histological images of murine stomach. Arrow, inflamed region. Scale bars, gross morphology, 1 cm; others, 50 um. a, antrum; c, corpus; d, duodenum; f, forestomach. (G) Histological gastritis score among WT, KO, and TKO mice (means +- SD, n = 4-5 mice/group). (H) IF for myeloperoxidase (MPO) in gastric antrum sections of 6-mo-old mice. Scale bars, 20 um. (I) Enumeration of MPO + cells in mucosa ( n = 4 mice/group). HPF, high-power field. (J) qPCR analysis of stomach lysates derived from 6-mo-old mice (means +- SD, n = 4-5 mice/group). Data are representative of two independent experiments. *, P < 0.05.

Supportive validation

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Experimental details: Figure 2 Hepatic Sirt6 is down-regulated during NASH development in mice. ( A ) H&E, Sirius Red staining, and immunofluorescence analysis of myeloperoxidase (MPO) and quantification of liver sections of control and WD-treated mice (4- to 6-month-old males, n = 4). ( D ) Western blot and quantification analysis of Sirt6, p-Smad2, Smad2, p-Smad3, and Smad3 in the livers of control and WD-treated mice. ( C ) Western blot and quantification of Smad3 acetylation (Ac-K-Smad3) in the livers of control and WD-treated mice. Data are presented as means +- SEM. * P < .05, ** P < .01, and *** P < .001 vs control; # P < .05, and ## P < .01 vs WD_4w.

Supportive validation

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Experimental details: Figure 6 Validation of CTSG, ELANE, and MPO protein levels. (A) Representative images of CTSG, ELANE, and MPO immunostaining in the lung. Original magnification, x 400. Scale bar represents 20 mum. (B) Quantification of CTSG, ELANE, and MPO immunostaining. Values are mean +- SD ( n = 5 for each treatment group. ** P < 0.01, *** P < 0.001 vs. model group; one-way ANOVA with Newman-Keuls multiple comparisons test).

Supportive validation

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Experimental details: Figure 2 NET formation induced by Toxoplasma gondii tachyzoites. Neutrophils were incubated for 180 min with RH strain tachyzoites (5:1 parasites:neutrophil ratio), fixed and stained for myeloperoxidase (MPO) and histone H1 ( A ) or T. gondii ( B ). Neutrophils DNA was counterstained with DAPI. Inset shows a region containing several parasites entrapped in DNA-MPO rich structures (arrows). Tg = T. gondii .