The tissue isolated from WT/Chow, WT/HF, MARCKS−/−/HF and MARCKS−/−/Chow groups of mice were fixed with 10% buffered formalin, imbedded in paraffin and sliced into 4–5 µm thick sections. Following hematoxylin and eosin (H&E) staining, the pathological changes of the liver tissues were observed under a light microscope. Hepatic lipid content in mice from WT/Chow, WT/HF, MARCKS−/−/HF, MARCKS−/−/Chow, CAL, and CAH groups was determined using 5-µm-thick frozen sections stained with Oil Red O (Sigma-Aldrich). Masson's trichrome staining was performed according to the manufacturer's instructions (Diagnostic Biosystems Inc., Pleasanton, CA, USA). In addition, some tissues also were subjected to immunohistochemical (IHC) staining for the analysis of MARCKS (except for MARCKS−/−/HF and MARCKS−/−/Chow groups) and PPARα from each group of mice expression. The sections were stained with MARCKS (ab51100; Abcam, Shanghai, China) and PPARα (ab8934; Abcam). All histological examinations were carried out according to standard procedures reported previously (27 (link)). Furthermore, immunofluorescence assays for PPARα for liver tissue were performed according to the manufacturer's instructions.
Ab51100
Manufactured by Abcam
Sourced in China
Ab51100 is a lab equipment product manufactured by Abcam. It is designed for use in scientific research and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (2 mentions)
Polyvinylidene fluoride membrane, by Merck Group (2 mentions)
Ab8934, by Abcam (1 mentions)
Oil red o, by Merck Group (1 mentions)
Masson s trichrome staining, by Diagnostic BioSystems (1 mentions)
Sc 180, by Santa Cruz Biotechnology (1 mentions)
Clone ac 74, by Merck Group (1 mentions)
Westernbright ecl hrp substrate, by Advansta (1 mentions)
Imagequant las 4000 scanner, by GE Healthcare (1 mentions)
Las 4000, by Cytiva (1 mentions)
3 protocols using ab51100
1
Histological Analysis of Liver Tissues
2
Quantifying Immune Signaling Proteins
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BMDM cell lysates were obtained following infection with L. monocytogenes or from untreated samples. BMDMs were infected at MOI 100:1 for 15 min, and the media was then replaced with DMEM containing gentamicin (100 mg/ml) for a further 105 min. Samples were clarified, denatured with SDS loading buffer, and boiled for 5 min. A total of 40 mg protein lysate was fractionated on 12% SDS-PAGE, transferred to polyvinylidene fluoride membranes (Millipore) and probed with primary Abs to murine MARCKS (1:2,000 dilution, ab51100, Abcam), murine RhoB (1:1,000 dilution, sc-180, Santa Cruz) or murine β-actin (1:10,000 dilution, Clone AC-74, Sigma), incubated with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology) and visualized using WesternBright ECL HRP substrate (Advansta), BD ChemiDoc system and ImageLab software.
3
Immunoblotting Analysis of BMDM Infection
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BMDM cell lysates were obtained following infection with E. coli NCTC12900 or from untreated samples. BMDMs were infected at MOI 20:1 for 15 min, and the media was then replaced with DMEM containing gentamicin (100 mg/ml) for a further 15 min. Samples were clarified, denatured with SDS loading buffer, and boiled for 5 minutes. A total of 20 mg protein lysate was fractionated on 12% SDS-PAGE, transferred to polyvinylidene fluoride membranes (Millipore) and probed with primary antibodies to murine MARCKS (1:2,000 dilution, ab51100, Abcam) (Lejonklou et al., 2012 (link)), CDC42 (1:500 dilution, 10155-1-AP, Proteintech), iNOS (1:500 dilution, 13120, Cell Signaling Technology) or β-actin (1:10,000 dilution, A3854, 3Sigma), incubated with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology) and visualized using Pierce™ ECL HRP substrate (Thermofisher) and ImageQuant LAS 4000 scanner (GE Healthcare).
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