Dulbecco's modified Eagle's medium (DMEM) was purchased from Gibco (Logan, UT, USA). Fetal bovine serum (FBS) was purchased from Yeasen Biotechnology Co., Ltd. Cell counting kit 8 (CCK8) was purchased from DOJINDO (Japan). Antibodies against cleaved‐caspase 3, p‐AMPK, p‐S6, p‐P70S6K, and SIRT1 were purchased from Cell Signaling Technology (USA); anti‐AMPK and β‐actin were purchased from Santa Cruz Biotech (USA). Antibody against PARP‐2 was purchased from Abcam (UK). Nobiletin was purchased from Chengdu Must Bio‐Technology Co., Ltd (Sichuan, China) and dissolved in dimethyl sulfoxide and stored at −20°C until diluted upon use. Recombinant adenovirus vectors expressing green fluorescent protein (Ad‐Flag) and Flag‐tagged PARP‐2 (Ad‐PARP‐2) were purchased from Genechem Co., Ltd. (Shanghai, China). The viruses were extended in HEK293A cells and purified by virus purification kit (Biomiga, USA), then dialyzed in dilution buffer and stored at −80°C.
Anti ampk
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-AMPK is a laboratory reagent that targets the AMP-activated protein kinase (AMPK) enzyme. AMPK is a key cellular energy sensor that plays a crucial role in regulating metabolism and cellular homeostasis. Anti-AMPK is designed to detect and study the AMPK enzyme in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Santa Cruz Biotechnology (6 mentions)
Anti p ampk, by Santa Cruz Biotechnology (4 mentions)
Anti sirt1, by Santa Cruz Biotechnology (3 mentions)
Anti p ampk, by Abcam (2 mentions)
Anti p62, by Santa Cruz Biotechnology (2 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
Anti lc3, by Merck Group (2 mentions)
Anti akt, by Santa Cruz Biotechnology (2 mentions)
Aicar, by Merck Group (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Penicillin, by Merck Group (2 mentions)
Ro 32 0432, by Merck Group (2 mentions)
Anti phospho akt, by Santa Cruz Biotechnology (2 mentions)
Anti lox 1, by Santa Cruz Biotechnology (2 mentions)
Anti ampkα, by Santa Cruz Biotechnology (2 mentions)
Anti β actin, by Cell Signaling Technology (2 mentions)
Anti phospho ampk, by Santa Cruz Biotechnology (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Yeasen (1 mentions)
19 protocols using anti ampk
1
Nobiletin Modulates Cellular Metabolism
2
Protein expression analysis of AMPK and stress response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Myocardial tissue samples were harvested, and total protein was extracted for electrophoresis and transferred to polyvinylidene difluoride (PVDF) membrane (Millipore, Burlington, MA, USA). The specific antibodies including anti-AMPK (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p-AMPK (1:200, Abcam, UK), anti-p38, anti-p-p38 (1:1,000, Affinity Company, San Francisco, CA, USA), anti-Nrf2 (1:1,000, Proteintech, Wuhan, China), and anti-HO-1 (1:1,000, Novus, Centennial, CO, USA) were added, and the mixture was incubated at 4 ℃ overnight. After washing 3 times with TBST, the secondary antibody (1:10,000, Santa Cruz Company, USA) was added. Blots were developed with electrochemiluminescence (ECL) substrate (Thermo Fisher, Waltham, MA, USA). Gray values were calculated using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
3
Cytoplasmic Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytoplasmic proteins of liver tissues and hepatocytes were extracted according to the instructions of the cytoplasmic protein extraction kit (Beyotime, Beijing, China). A total of 30–60 μg cytoplasmic protein was resolved on an 8%–12% precast gel using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (Invitrogen, NY, USA) and transferred to polyvinylidene fluoride membranes (Bio-Rad, CA, USA). The membranes were then incubated with anti-p-AMPK, anti-AMPK, anti-G6Pase, anti-PEPCK, anti-ACC, anti-p-ACC, anti-FAS, anti-CPT-1a, anti-SHP1, anti-SREBP-1c (Santa Cruz, CA, USA), and anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Cell Signaling, MA, USA). Then, these membranes were washed with PBS and incubated with anti-mouse or anti-rabbit secondary antibody (Beyotime) for 1 h at room temperature. Finally, the immune complexes were developed using an enhanced chemiluminescence western blotting substrate, and protein expression levels were quantified using ImageJ software.
4
Adipogenesis Regulation via Autophagy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Icariin, L-asparagine (Asn), 3-methyladenine (3-MA), Oil Red O, MTT assay kit, uranyl acetate/lead citrate, and MDC were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin, adipocyte differentiation medium, and gentamycin were purchased from Hyclone Laboratories, Inc. (Logan, UT, USA). The Annexin V-FITC apoptosis detection kit was purchased from BD Biosciences (Franklin Lakes, NJ, USA). Anti-mTOR, anti-p-mTOR, anti-beclin-1, anti-AMPK, anti-p-AMPK, anti-p62, anti-LC3, and anti-β-actin antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
5
Regulation of Autophagy by FOXO1 and AMPK
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NaB and SAHA were purchased from Sigma Aldrich (Deisenhofen, Germany). The FOXO1 inhibitor AS1842856 (AS) was obtained from Selleck, and the adenosine monophosphate-activated protein kinase (AMPK) inhibitor was purchased from MCE. The monoclonal anti-ubiquitin, anti-acetylation, anti-Snail, anti-Smad2, anti-Smad3, anti-Smad2/3, anti-p-Smad2/3, anti-FOXO1, anti-LC3B, anti-P62, anti-ULK1, anti-p-ULK1, anti-LC3, anti-GAPDH, and anti-vimentin antibodies and the secondary anti-mouse or anti-rabbit antibody conjugated to horseradish peroxidase (HRP) were acquired from Cell Signaling Technology (MA, USA). The secondary anti-rabbit antibody conjugated to fluorescein isothiocyanate (FITC), Protein A/G Sepharose and anti-AMPK were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p-AMPK and anti-FN were acquired from Abcam. A Millicell chamber (8 mm) was purchased from Millipore (BD Biosciences, USA). The Annexin V, FITC apoptosis detection Kit S was obtained from Japan Tongren Chemical. SYBR Premix ExTaq II was a product of TaKaRa (TBI, Japan). siRNA FOXO1 and siRNA ULK1 were purchased from RIBO, and Lipofectamine 3000 was purchased from Invitrogen (Carlsbad, CA, USA). 4,6-diamidino-2-phenylindole (DAPI) dye was obtained from FUDE company.
6
Western Blot Analysis of Autophagy Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cell extracts preparation and Western blot analysis were performed as previously described [13 (link)]. The antibodies used were anti-BRAF and anti-AMPK (Santa Cruz Biotechnology, Dallas, TX, USA); anti-p62 (BD Biosciences, Franklin Lakes, NJ, USA); anti-ATG5, anti-phospho-LKB1 (S428), anti-LKB1, anti-phospho-ACC (S79), anti-phospho-AMPK (T172), anti-phosho-S6 (S235/236) and anti-phospho-ULK (S555) (Cell Signalling Technology, Danvers, MA, USA); anti-LC3, anti-phospho-ERK and anti-β-tubulin (Sigma, St. Louis, MO, USA); and peroxidase-conjugated secondary antibodies (DAKO, Glostrup, Denmark). For LC3-II/LC3-I ratio calculation, data densitometry of blots from different experiments was used to quantify the protein bands.
7
Western Blot Analysis of Metabolic Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues or cells were lysed in RIPA buffer (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 12 mM 464 deoxycholic acid, 0.5% Nonidet P-40, and protease and phosphatase inhibitors). Ten μg proteins were loaded on SDS-PAGE and subjected to immunoblotting. Nitrocellulose membranes were incubated with anti-Ampk-p172 (#2531, Cell Signaling Technology), anti-Ucp1 (ab23841, abcam), anti-vDAC (sc-32063, Santa Cruz Biotechnology), anti-Hsp60 (sc-13966, Santa Cruz Biotechnology), anti-Ampk (sc-25792, Santa Cruz Biotechnology), anti-Hsl (#4107, Cell Signaling Technology), Pc (sc-271493, Santa Cruz Biotechnology), anti-Tom20 (sc-11415, Santa Cruz Biotechnolog), anti- Complex I ndufb8 (ms-105, MitoSciences), anti-Complex III Core2 subunit (ms-304, MitoSciences), anti-Vinculin (ab-18058, Abcam), and anti-Pdhβ (gtx-119625, GeneTex) primary antibodies, at 1:1000 dilution. Successively, membranes were incubated with the appropriate horseradish peroxidase-conjugated secondary antibodies. Immunoreactive bands were detected with a FluorChem FC3 System (Protein-Simple, San Jose, CA, USA) after incubation of the membranes with ECL Selected Western Blotting Detection Reagent (GE Healthcare, Pittsburgh, PA, USA). Densitometric analyses of the immunoreactive bands were performed with the FluorChem FC3 Analysis Software.
8
Synthetic Soya-Cerebroside Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soya-cerebrosides (Fig. 1A ) of >98% purity were synthesized at the College of Pharmacy, School of Pharmacy, China Medical University, Taichung, Taiwan (The detail synthesis procedures and purified quality was provided as described previously21 (link)). Soya-cerebroside was dissolved in DMSO for intraperitoneal (IP) injection. A miR-432 inhibitor, Control miRNA and TRIzol were from Life Technologies (Carlsbad, CA). Rabbit polyclonal antibody specific for MCP-1, MCP-1 enzyme-linked immunosorbent assay (ELISA) kit and human recombinant IL-1β were purchased from R&D Systems (Minneapolis, MN, USA), anti-phospho AMPK (Thr172) and phospho AKT (Ser473) from Cell Signaling (Danvers, MA, USA). Mouse monoclonal CD68 was purchased from Novus (Novus, Littleton, USA) and Ara A (ATP mimetic), Compound C (AMPK kinase inhibitor), and AKTi (AKT1/2 kinase inhibitor) from Enzo Biochem, Inc. (Enzo, New York, NY). Anti-AMPK, anti-AKT, and anti-mouse and anti-rabbit conjugated horseradish peroxidase (HRP) antibodies were from Santa Cruz (Santa Cruz, CA, USA). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).
9
Western Blotting of Hippocampal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described [27 (link)]. In brief, the frozen right hippocampus was lysed in RIPA buffer containing 150 mM NaF, 2 mM sodium orthovanadate, and protease inhibitors (protease inhibitor mixture; Roche). Protein of total lysate (20 μg) was loaded and blotted. The membranes were incubated with primary antibodies anti-IDE (MMS-282R; 1:1000; Covance, U.K.), anti-PPARγ (81B8, Cell Signaling Technology, Beverly, MA, U.S.A.), and anti-AMPK (1:1000; Santa Cruz Biotechnology, CA, U.S.A.) overnight at 4˚C, and then reacted with HRP-conjugated secondary antibodies (1:1000; Santa Cruz Company) at room temperature for 1.5 h. The protein bands were detected by ECL and visualized by UVP Gel imaging system (Upland, CA). The band intensity was analyzed by AlphaEaseFC (version 4.0).
10
HUVEC Culture and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human umbilical vein endothelial cells (HUVECs) were obtained from ATCC. HUVECs were cultured with M199 basal medium supplemented with low-serum growth supplement and penicillin (50 IU/ml)-streptomycin (50 μg/ml). trypsin-EDTA was used to passage cells. M199 and trypsin-EDTA were obtained from Gibco (Grand Island, NY, USA). Low-serum growth supplement was purchased from Cascade (Portland, OR, USA). Additionally, 5,58,6,68-tetraethylbenzimidazolcarbocyanine iodide (JC-1) were purchased from BioVision (Palo Alto, CA, USA). LY294002, dihydroethidium (DHE), Apocynin, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), DCHC, AICAR, sodium nitroprusside (SNP), Ro-320432, penicillin and streptomycin were all purchased from Sigma (St. Louis, MO, USA). Anti-β-actin, anti-AMPK, anti-AMPK-α, anti-AKT, anti-phospho AKT, anti-LOX-1, anti-SIRT1, anti-eNOS were all obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). HRP-conjugated anti-rabbit secondary antibodies were purchased from Transduction Laboratories (CA, USA). (TUNEL) staining kit was obtained from Boehringer Mannheim (Mannheim, Germany) was purchased from Selleckchem. Fura-2 AM and the EnzChek caspase 3 assay kit were purchased from Molecular Probes (Eugene, OR). SOD activity assay kit was obtained from Calbiochem (San Diego, CA). eNOS kit was purchased from R&D Systems (Minneapolis, MN, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!