Liver was lysed in RIPA buffer supplemented with Halt Protease and Phosphatase Inhibitor Cocktail (Thermo-Fisher, Waltham, MA) and 1 mM PMSF. Protein concentrations were determined using Pierce BCA Protein Assay Kit (Pierce, Rockford, IL). Twenty-five micrograms of protein per lane were loaded onto a 4–12% Criterion TGX Precast Gel (Bio-Rad, Hercules, CA) then transferred to PVDF membranes using a Trans-Blot Turbo Transfer System (Bio-Rad, Hercules, CA). Membranes were blocked in 5% non-fat milk followed by overnight incubation with primary antibody at 4 °C. Primary antibodies used are as follows: anti-ACADM (55210, Proteintech, Rosemont, IL) and anti-CPT2 (ab181114, Abcam, Cambridge, MA). Following primary antibody incubation, the blots were washed and incubated with an anti-rabbit IgG HRP-conjugated secondary antibody (#7074S, Cell Signaling Technology, Danvers, MA). Blots were stripped with Restore Western Blotting Stripping Buffer (Thermo-Fisher, Waltham, MA) and re-probed with anti-ACTB for normalization (ab8227, Abcam, Cambridge, MA). Blot imaging was performed on a ChemiDoc™ MP System (Bio-Rad, Hercules, CA) after exposing blot to Clarity Western ECL Blotting Substrate (Bio-Rad, Hercules, CA).
Anti acadm
Manufactured by Proteintech
Sourced in China
Anti-ACADM is a primary antibody that recognizes the ACADM protein. ACADM, also known as medium-chain acyl-CoA dehydrogenase, is an enzyme involved in the breakdown of medium-chain fatty acids. This antibody is commonly used in various research applications, such as Western blotting and immunohistochemistry, to detect and study the ACADM protein.
Automatically generated - may contain errors
Lab products found in correlation
Restore western blotting stripping buffer, by Thermo Fisher Scientific (1 mentions)
Ab181114, by Abcam (1 mentions)
Clarity western ecl blotting substrate, by Bio-Rad (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Hrp conjugated anti rabbit igg secondary antibody, by Cell Signaling Technology (1 mentions)
Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Chemidoc mp system, by Bio-Rad (1 mentions)
Ab8227, by Abcam (1 mentions)
Anti acox1, by Proteintech (1 mentions)
Anti acaa1, by Proteintech (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Bca protein assay reagent kit, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ecl reagent, by Merck Group (1 mentions)
Anti bcl 2 antibody, by Abcam (1 mentions)
Anti hadh, by Proteintech (1 mentions)
Anti glut1, by Abcam (1 mentions)
Peg300, by Merck Group (1 mentions)
Filipin 3, by Santa Cruz Biotechnology (1 mentions)
Diethylnitrosamine, by Tokyo Chemical Industry (1 mentions)
3 protocols using anti acadm
1
Protein Expression Analysis in Liver Tissue
2
Western Blot Analysis of Metabolic Proteins
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Tumor or liver tissues and cells were lysed by 1% SDS lysis buffer and boiled at 95 °C for 10 min, then centrifuged at 12,000 rpm for 10 min at room temperature. The protein concentration was determined by a BCA protein assay reagent kit (Thermo Scientific, Waltham, MA, USA). The protein (35 μg) was separated by SDS-PAGE gels and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked for 2h by 5% fat free milk in TBST at room temperature, followed by incubation overnight at 4 °C with the primary antibodies for anti-Glut1 (Abcam, Cambridge, UK), anti-Bcl2 antibody (Abcam), anti-Pparg antibody (Proteintech, Wuhan, Hubei, China), anti-Acaa1 (Proteintech), anti-Acadm (Proteintech,), anti-CD36 (Bioworld, Nanjing, China), anti-Hadh (Proteintech), anti-RXRA (Proteintech), anti-ACOX1 (Proteintech), and anti-β-actin (Sigma), respectively. The membranes were washed with TBST and incubated for 1 h with the corresponding HRP-conjugated second antibodies. Bands were visualized with ECL Reagents (Merck, Billerica, MA, USA).
3
Lipid Metabolism Regulation in Liver Disease
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Triton X-100, PEG300, and Tween 80 were purchased from Sigma-Aldrich. Bodipy 493/503 was purchased from Thermo Fisher Scientific. Filipin III was purchased from Santa Cruz. Etomoxir, avasimibe, palmitic acid, and oleic acid were purchased from Tsbiochem in China. Diethylnitrosamine was purchased from Tokyo Chemical Industry in Japan. 60% HFD was purchased from Moldiets, Biopike in China. Rabbit anti-CDK4(ab108357, 1:1000 for WB), rabbit anti-CDK6 (b124821, 1:1000 for WB) were purchased from Abcam (Cambridge, UK). Rabbit anti- SOAT1 (A6311, 1:1000 for WB) was purchased from Abclonal. Rabbit anti-SOAT1 (NB400-141, 1:1000 for IHC) was purchased from Novus. Rabbit anti-CyclinD1 (26939-1-AP, 1:1000 for WB, 1:100 for IHC), Anti-CPT1A (15184-1-AP, WB: 1/1000; IHC:1/100), Anti-Perilipin 3/TIP47 (10694-1-AP, WB: 1/1000; IHC 1/100), Anti-ACADM (55210-1-AP, WB: 1/500), Anti-ACADL (17526-1-AP, WB: 1/500), Anti-GAPDH were purchased from Proteintech.
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