Immunostaining of frozen cross sections used a rat monoclonal anti-mouse Ly6G (clone 1A8), rabbit polyclonal anti-citrullinated Histone H3 (H3cit, Abcam) and anti-citrullinated Histone H4 (H4cit, Millipore), a rat monoclonal anti-CD31 (BD Pharmingen), a rabbit polyclonal anti-CD66b (Abcam) or a rabbit polyclonal anti-myeloperoxidase (DAKO). Immunostaining was performed on human coronary artery endothelial cells (HCAEC) using a rabbit anti-C5b-9 complement (Abcam) or a rabbit anti-VE-Cadherin (Abcam). Apoptotic ECs were visualized using fluorescent in situ DNA strand breaks detection kit (TUNEL, Roche). Immunostaining was amplified using peroxidase-conjugated streptavidin complexes (Vector Laboratories) and peroxidase was detected using AEC (Vector Laboratories) substrate. Sections were lightly counterstained with hematoxylin, mounted in gelatin-glycerol and examined with a bright field microscope (Nikon Optiphot-2 equipped with a Nikon digital camera DXM 1200F). For double immunostaining studies, cross sections were incubated with primary antibodies followed by incubation with fluorophore-coupled anti-species antibody (Life Technologies), stained with DAPI, and mounted with fluorescent mounting medium (DAKO). Slides were kept in the dark at 4°C.
Anti myeloperoxidase
Manufactured by Agilent Technologies
Sourced in United States
Anti-myeloperoxidase is a lab equipment product used to detect and measure the levels of myeloperoxidase, an enzyme found in certain white blood cells. It serves as a tool for diagnostic and research purposes, providing information about the presence and concentration of myeloperoxidase in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd31, by BD (1 mentions)
Optiphot 2, by Nikon (1 mentions)
Dxm1200f, by Nikon (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Anti cd66b, by Abcam (1 mentions)
Rabbit anti ve cadherin, by Abcam (1 mentions)
Lsm 780 confocal microscope, by Zeiss (1 mentions)
Prolong gold antifade, by Thermo Fisher Scientific (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Alexa 568, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Evolution mp camera, by Media Cybernetics (1 mentions)
Axiocam mrm, by Zeiss (1 mentions)
Axiovision software, by Zeiss (1 mentions)
Axiovert 200, by Zeiss (1 mentions)
4 protocols using anti myeloperoxidase
1
Immunostaining of Coronary Artery Endothelial Cells
2
Neutrophil NET formation imaging
3
Neutrophil Immunofluorescence Staining
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Untreated, PMA or Candida ssp. treated neutrophils (2 x 105 per condition) were fixed with 4% paraformaldehyde, permeabilized (0.5% triton X-100) for 15 minutes and blocked with 3% BSA. After washing, the cells were stained for 1 h at 37 °C with anti-myeloperoxidase (1:400) (Dako, Agilent Technologies, Carpinteria, USA) and anti-calprotectin (clone MAC 387; 1:400) (abcam). Two more washes were made, followed by addition of secondary antibodies coupled to Alexa 568 and Alexa 488 (Molecular Probes) and incubated for 1 h at 37 °C. Next, cells were counterstained with ProLong Gold antifade reagent with DAPI (P36941; Molecular Probes). Epifluorescence images were captured using a Zeiss Axioplan upright epifluorescence microscope with 100× objectives and images were captured using a Media Cybernetics Evolution MP camera and Q Imaging software.
4
Immunofluorescence Imaging of Cardiac Tissue
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Perfused hearts were snap frozen in optimal cutting temperature medium and cryosectioned at 8‐ to 10‐μm thickness. After fixation with 2% paraformaldehyde, sections were blocked in 3% bovine serum albumin, and then incubated with anti‐CD41 (clone MWReg30; 0.5 μg/mL; Biolegend) or anti‐myeloperoxidase (0.32 μg/mL; Dako) antibodies overnight at 4°C. After washing, samples were incubated with Alexa555‐conjugated anti‐rat IgG or Alexa488‐conjugated anti‐rabbit IgG (1.5 μg/mL) for 2 hours at RT. Sections were then washed, and DNA was counterstained with Hoechst 33342 (1:10 000) for 10 minutes before mounting with antifade Fluorogel mounting medium (Electron Microscopy Sciences). Images were acquired using a Zeiss Axiovert 200 inverted wide‐field fluorescence microscope coupled to an Axiocam MRm monochromatic charge‐coupled device camera. Channels were acquired in grayscale and pseudocolored with Zeiss Axiovision software.
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