Smooth muscle cells were stained with anti-α-smooth muscle actin (SMA; ab5694, Abcam), which detects smooth muscle α-actin 2 (ACTA2), and macrophages were stained using macrophages/monocytes antibody (MOMA-2; MCA519G, Bio-Rad). Slides were fixed in cold acetone at −20°C and air dried, and endogenous peroxidase activity was blocked in 0.3% H2O2/methanol. Nonspecific binding was reduced by incubation in 2.5% goat serum (Vector Laboratories) before primary antibody incubation at 4°C in a humidified chamber overnight. Slides were incubated with ImmPRESS HRP anti-rabbit (Vector Laboratories, MP-7451) or anti-rat IgG peroxidase (Vector Laboratories, MP-7444), and staining was visualized using DAB and counterstained in hematoxylin (Gill No. 2, Sigma-Aldrich). Images were acquired with a DM2500 Leica microscope. Positively stained areas of each aorta were measured with ImageJ by a researcher blinded to genotype. The data are presented as the average of positively stained lesion areas (9 serial sections) normalized by total lesion area. For collagen and lipid core quantification, sections were stained with Masson’s trichrome (Sigma-Aldrich) according to the manufacturer’s instructions.
Mca519g
Manufactured by Bio-Rad
Sourced in United States
The MCA519G is a laboratory equipment product manufactured by Bio-Rad. It is a sensitive and accurate instrument designed to perform specific tasks in a research or diagnostic setting. The core function of this product is to facilitate precise measurements and analyses, but a detailed description of its intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Goat serum, by Vector Laboratories (1 mentions)
Gill no 2, by Merck Group (1 mentions)
Leica dm2500 microscope, by Leica camera (1 mentions)
Immpress hrp anti rabbit, by Vector Laboratories (1 mentions)
Masson s trichrome, by Merck Group (1 mentions)
Ab5694, by Abcam (1 mentions)
D9542, by Merck Group (1 mentions)
D7008, by Merck Group (1 mentions)
2 protocols using mca519g
1
Quantifying Aortic Lesions and Components
2
Evaluating Oxidative Stress and Atherosclerosis in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After 13 weeks on Western diet, mice were euthanized and tissues were harvested. For ROS staining, cryosections of liver were incubated with fluorescent dye DHE (D7008, Sigma-Aldrich) and the nuclei were counterstained with DAPI (D9542, Sigma-Aldrich). Then, green, red, and blue fluorescence were imaged using a fluorescence microscope, respectively. The ROS levels of liver sections were calculated using Image-Pro Plus 6.0 and represented by the mean DHE fluorescence intensity in the detected area. For analysis of the atherosclerotic lesions in aorta, 8-μm serial sections of the aortic root and en face of the aorta were examined by Oil Red O staining, 27 and lesions were quantified with Image-Pro Plus 6.0. Aortic lesion area is represented as a percentage of the total aorta area, and the atherosclerotic plaque of aortic root is represented as mean lesion area (μm 2 ). Macrophages on sections of the aortic root were determined by immunohistochemical analysis using rat anti-mouse MOMA2 antibody (MCA519G, Bio-Rad, Hercules, CA, USA). Histochemical analysis for liver steatosis was performed by hematoxylin and eosin (H&E) staining and Oil Red O staining.
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