The following materials were obtained from Sigma-Aldrich (St. Louis, MO, USA): 5-HT, N-(3-dimethylaminopropyl)-Nʹ-ethylcarbodiimide hydrochloride (EDC), and N-hydroxysuccinimide (NHS), lactoperoxidase (LPO). Fe3O4@PEG-COOH was purchased from Nanjing Nanoeast Biotech Co., Ltd. (Nanjing, China). Vivotrax and Perimag were purchased from Magnetic Insight Inc (Alameda, USA) and Micromod Partikeltechnologie GmbH (Rostock, Germany), respectively. Cy7-NHS was obtained from AAT Bioquest (Sunnyvale, CA, USA). RAW 264.7 cells (Otwo Biotech Inc., Shenzhen, China), Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), fetal bovine serum (FBS; Gibco, Grand Island, NY, USA), and cell counting kit-8 (CCK-8; Beyotime Institute of Biotechnology, Shanghai, China) were obtained for cell experiments. Myeloperoxidase (Abcam, Cambridge, UK), glucose oxidase (Dalian Meilun Biotechnology Co., Ltd., Dalian, China), Matrigel (Corning, Inc., Corning, NY, USA), 4-ABAH (Sigma-Aldrich), and ExiTron (TM) nano 12000 CT contrast agent (Miltenyi Biotec, Bergisch Gladbach, Germany) were purchased for animal experiments. Erythropoietin protein (EPO), rabbit anti-mouse MPO, rabbit anti-mouse CD68, rabbit anti-mouse anti-ɑ-smooth muscle actin (SMA), rabbit anti-mouse CD31, and rabbit anti-mouse MCP-1 antibodies were obtained from Abcam.
Rabbit anti mouse cd68
Manufactured by Abcam
Sourced in United Kingdom, Switzerland
Rabbit anti-mouse CD68 is a primary antibody that detects the CD68 protein, a heavily glycosylated transmembrane protein expressed predominantly by cells of the monocyte-macrophage lineage in mice.
Automatically generated - may contain errors
Lab products found in correlation
Cy7 nhs, by AAT Bioquest (1 mentions)
N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride edc, by Merck Group (1 mentions)
4 abah, by Merck Group (1 mentions)
Rabbit anti mouse cd31, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Matrigel, by Corning (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
E400 microscope, by Nikon (1 mentions)
Histochoice, by Avantor (1 mentions)
3 protocols using rabbit anti mouse cd68
1
Multimodal Imaging Probe Characterization
2
Immunohistochemical Analysis of Inflammatory Markers
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The paraffin tissue sections were dewaxed and then treated with 0.3% hydrogen peroxide solution for 10 min. Then, the sections were immersed in 0.01 mol/L citric acid buffer and heated to boiling in an autoclave 3 min 3 times. Next, the sections were blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature. Then, the samples were washed three times with phosphate-buffered saline (PBS) and incubated with primary antibodies (rat anti-mouse α-actin [dilution 1:100], rabbit anti-mouse IFN-γ [dilution 1:100] and rabbit anti-mouse CD68 [dilution 1:50], all purchased from Abcam, United Kingdom) for 1 h at 37°C. The sections were then rinsed three times with PBS, treated with appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies for 30 min at 37°C and washed 3 times with PBS again. Antigen-antibody reactions were stained with diaminobenzidine (DAB) for 5–10 min, and all sections were counterstained with hematoxylin. The expression levels of CD68, IFN-γ and α-actin were observed with a Nikon E400 microscope under high-power (400×) fields.
3
Aortic Plaque Quantification and Characterization
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En face preparations of the aorta (descending and arch) were fixed in Histochoice (Amresco, Solon OH, USA) and stained with 0.16% Oil-Red-O dissolved in 78% methanol/0.2 mol/L NaOH.
Plaques were identified as Bordeaux colored regions and expressed as percentage out of total aorta area and quantified by one blinded observer. For analysis of aortic root plaques, frozen tissue sections were cut from the aortic root, frozen and stained using the following primary antibodies: rat anti-mouse IL-22RA1 antibody (R&D systems, Abingdon, UK), rabbit anti-mouse SMC α-actin antibody (Abcam, Cambridge, UK), rat anti-mouse MOMA-2 antibody (BMA Biomedicals, Augst, Switzerland), rabbit-anti-mouse Caldesmon (Abcam), rabbit-anti-mouse CD68 (Abcam). Stainings were developed using ABC-elite DAB detection kit, according to manufacturer's instructions (Vector Laboratories, Burlingame, CA, USA) and counterstained with Haematoxylin. Exclusion of the primary antibody or staining with IgG isotype control antibodies was used as controls. For analysis of collagen and lipid content in aortic root plaques, Van Gieson and Oil-Red-O stainings were used. Stained plaque areas were quantified blindly using BioPix iQ 2.0 software (Biopix AB, Gothenburg, Sweden). Aortic root plaque area was calculated as the mean of three separate sections across the aortic root region.
Plaques were identified as Bordeaux colored regions and expressed as percentage out of total aorta area and quantified by one blinded observer. For analysis of aortic root plaques, frozen tissue sections were cut from the aortic root, frozen and stained using the following primary antibodies: rat anti-mouse IL-22RA1 antibody (R&D systems, Abingdon, UK), rabbit anti-mouse SMC α-actin antibody (Abcam, Cambridge, UK), rat anti-mouse MOMA-2 antibody (BMA Biomedicals, Augst, Switzerland), rabbit-anti-mouse Caldesmon (Abcam), rabbit-anti-mouse CD68 (Abcam). Stainings were developed using ABC-elite DAB detection kit, according to manufacturer's instructions (Vector Laboratories, Burlingame, CA, USA) and counterstained with Haematoxylin. Exclusion of the primary antibody or staining with IgG isotype control antibodies was used as controls. For analysis of collagen and lipid content in aortic root plaques, Van Gieson and Oil-Red-O stainings were used. Stained plaque areas were quantified blindly using BioPix iQ 2.0 software (Biopix AB, Gothenburg, Sweden). Aortic root plaque area was calculated as the mean of three separate sections across the aortic root region.
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