For histology analysis, we imbedded the fixed livers in paraffin, then cut thin sections, and following standard procedures, stained them with hematoxylin and eosin. We quantified the liver histological score of inflammation according to Ishak inflammation score [43 (link), 44 (link)]. For immunofluorescence analysis, we cut frozen sections into 8μM slices in a cryostat microtome (Thermo) at -20°C and permeabilized and blocked them. We probed the tissues with appropriate, fluorescently labelled antibodies, including APC-conjugated anti-CD11b, PE-conjugated anti-CD68, and PE-conjugated anti-Ly6G (Biolegend). Finally, we dried the sections and observed them with a Nikon A1 confocal microscope.
Pe conjugated anti cd68
Manufactured by BioLegend
PE-conjugated anti-CD68 is a monoclonal antibody that binds to the CD68 antigen, which is a glycoprotein expressed on the lysosomal membrane of monocytes and macrophages. The antibody is conjugated with the fluorescent dye Phycoerythrin (PE) to enable detection and analysis of CD68-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cryostat microtome, by Thermo Fisher Scientific (1 mentions)
Apc conjugated anti cd11b, by BioLegend (1 mentions)
Pe conjugated anti ly6g, by BioLegend (1 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions)
Moflo astrios eq, by Beckman Coulter (1 mentions)
70 m cell strainer, by Corning (1 mentions)
Rat anti mouse cd16 cd32 antibody, by BD (1 mentions)
2 protocols using pe conjugated anti cd68
1
Histological and Immunofluorescence Analysis of Liver
2
Isolation and Characterization of Aortic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mice were euthanized with inhaled isoflurane overdose and the circulatory system was cleared by perfusion with PBS. Aortas were dissected and incubated with collagenase type II (175 U/ml) to remove adventitia. Aortas were minced and placed into an enzyme cocktail (collagenase type I 450 U/ml, collagenase type XI 125 U/ml, hyaluronidase 60 U/ml). Cell suspension was passed through a 70 µM cell strainer (Corning, Corning, NY) and washed with FACS buffer (PBS, 1% BSA, 1 mM EDTA). Samples were fixed and permeabilized with Foxp3/Transcription Factor Staining Buffer Set (Thermo Fisher). Cells were blocked with rat anti-mouse CD16/CD32 antibody (BD Biosciences) and stained with PE-conjugated anti-CD68 (BioLegend, San Diego, CA) and FITC-conjugated anti-α-smooth muscle actin (Sigma-Aldrich) antibodies for 45 min 4 °C. The samples were run on MoFlo Astrios EQ (Beckman Coulter, Indianapolis, IN) and data were analyzed with FlowJo v10 (FlowJo, LLC, Stanford University, CA).
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