Hearts were perfused through the left ventricle with 1X phosphate‐buffered saline (PBS; Thermo Scientific, Waltham, MA), followed by 4% paraformaldehyde (Sigma‐Aldrich) and cut and embedded in optimal cutting temperature (OCT) compound (Tissue‐Tek 4583). This was followed by frozen sectioning on a microtome‐cryostat (10 µm) starting from where the aorta exits the ventricle and moving toward the aortic sinus. Sections were stained with ORO or trichrome (Gomori's Trichrome Stain Kit, 87020; Thermo Scientific) or immunostained with monocyte and macrophage 2 (MOMA‐2) antibody (1:500; Abcam, Cambridge, United Kindom) overnight at 4°C before incubating with secondary antibody for 1 hour (immunohistochemistry‐labeled streptavidin biotin kit; Dako). Images were taken at 4× using an Olympus SZX7‐TR30 microscope and quantified with the CellSens Standard program.
Moma 2 antibody
Manufactured by Abcam
The MOMA-2 antibody is a research-use antibody that targets the MOMA-2 antigen. It is commonly used in flow cytometry and immunohistochemistry applications. The antibody is available in different formats, such as purified and conjugated versions, to suit various experimental requirements.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780, by Zeiss (2 mentions)
Gomori s trichrome stain kit, by Thermo Fisher Scientific (1 mentions)
Szx7 tr30 microscope, by Olympus (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Amylase antibody, by Santa Cruz Biotechnology (1 mentions)
Cd11b antibody, by Abcam (1 mentions)
Cerulein, by MedChemExpress (1 mentions)
Zm 241385, by Selleck Chemicals (1 mentions)
Gsk2606414, by Selleck Chemicals (1 mentions)
Abt 702, by Merck Group (1 mentions)
Grp78 antibody, by Proteintech (1 mentions)
Chop antibody, by Proteintech (1 mentions)
Perk antibody, by Proteintech (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Masson staining kit, by Solarbio (1 mentions)
Hur antibody, by Merck Group (1 mentions)
α sma antibody, by Abcam (1 mentions)
α sma antibody, by Merck Group (1 mentions)
4 protocols using moma 2 antibody
1
Histological Analysis of Aortic Tissues
2
Cerulein-Induced Pancreatitis Signaling
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Cerulein was purchased from MedChem Express (HY-A0190, Shanghai). Lipopolysaccharide (LPS) was purchased from Sigma (L2630, Shanghai). ABT702 was purchased from Merck Millipore (116890, Shanghai). ZM241385 (an adenosine A2A receptor antagonist) and GSK2606414 (PERK inhibitor) were purchased from Selleck (S8105 and S7307, respectively, Shanghai). The following primary antibodies were purchased: ß-actin antibody (Proteintech, 66009), ADK antibody (Abcam, ab227087), RIP1 antibody (CST, 3,493), phosphor-RIP1 (Ser161) antibody (Affinity, AF7377), RIP3 antibody (CST, 15828), phosphor-RIP3 antibody (Abcam, ab195117), mouse MLKL antibody (Proteintech, 66675), rat MLKL antibody (Abcam, ab243142), phosphor-MLKL antibody (Affinity, AF7420), NF-κB p65 antibody (CST, 8242), phosphor-NF-κB p65 antibody (CST, 3,033), GRP78 Antibody (Proteintech, 11587), CHOP antibody (Proteintech, 15204), PERK antibody (Proteintech, 24390), phosphor-PERK antibody (CST, 3,179), eIF2α antibody (CST, 5324), phosphor-eIF2α antibody (CST, 3,398), CD11b antibody (Abcam, ab133357), MOMA-2 antibody (Abcam, ab33451), and amylase antibody (Santa Cruz Biotechnology, sc-46657). Secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, United States).
3
Quantifying Aortic Plaque Vulnerability
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Aortas and hearts were embedded in OCT and sliced into 5-μm-thick frozen sections. The heart was cut to the aortic root. After sections were hydrated, immunofluorescence, immunohistochemistry, or other special staining was performed. To detect HuR-knockout efficiency in VSMCs, fluorescent double labeling of aortic sections was performed with HuR antibody (1:300, Millipore) and α-SMA antibody (1:300, Abcam). Sections of aortic roots were immunostained with MOMA-2 antibody (1:300, Abcam) to detect macrophage content and α-SMA antibody to detect VSMC content. The collagen of aortic root was detected by using a Masson staining kit (Solarbio, Beijing). The plaque vulnerability index was calculated as follows: (macrophage staining % + lipid staining %)/(SMC staining % + collagen staining %)53 (link).
4
Immunofluorescence Analysis of Aortic Macrophages
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The frozen sections of aortic root were fixed with ice-acetone for 10 min, added with 0.3% Triton X-100 to make cell membrane permeabilized, and blocked with 5% BSA at room temperature for 1 h. Then they were blocked at 4°C overnight with primary antibodies (MOMA-2 antibody 1 : 50, Abcam; α-SMA antibody 1 : 400, Sigma), and incubated at room temperature for 3 h using secondary antibodies (1 : 200 and 1 : 400) protecting from light for coloration by antigen-antibody binding. Finally they were counterstained for nuclei with DAPI and mounted with the fluorescer. Macrophages and smooth muscle actin were observed under confocal fluorescence microscope (Zeiss LSM 780) and the positively stained macrophages and smooth muscle cells were automatically analyzed and quantified on Image-pro plus 6.0.
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