FC (purity is 99.5% by high performance liquid chromatography) is obtained from endophytic Aspergillus terreus (strain No. FC118) by the root of Rhizophora stylosa (Rhizophoraceae). The primary antibodies including PPAR-α, PPAR-β, PPAR-γ, C/EBP-α, C/EBP-β, SREBP-1c, aP2, LPL, FAS, HSL, AQP-7, ATGL, β-actin, and alkaline phosphatase labeled secondary antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). Glycerol, TG, total cholesterol (TC), and Cell Counting Kit-8 (CCK-8) were evaluated via diagnostic assay kits from Nanjing Jiancheng Company (Nanjing, China). Simvastatin (Sim) was purchased from Sigma Aldrich Company (St. Louis, MO, USA).
Pparα
Manufactured by Cell Signaling Technology
Sourced in United States
PPARα is a nuclear receptor that functions as a ligand-activated transcription factor. It regulates the expression of genes involved in energy homeostasis, lipid metabolism, and inflammation.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Cell Signaling Technology (5 mentions)
Srebp 1c, by Cell Signaling Technology (4 mentions)
Srebp1, by Cell Signaling Technology (4 mentions)
Pparγ, by Cell Signaling Technology (3 mentions)
C ebpα, by Cell Signaling Technology (3 mentions)
Pgc 1α, by Cell Signaling Technology (3 mentions)
Bca kit, by Thermo Fisher Scientific (3 mentions)
Cpt1α, by Cell Signaling Technology (2 mentions)
Cpt1a, by Cell Signaling Technology (2 mentions)
P ampk, by Cell Signaling Technology (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
Nf κb p65, by Cell Signaling Technology (2 mentions)
Bicinchoninic acid analysis, by Thermo Fisher Scientific (2 mentions)
P s6k, by Cell Signaling Technology (2 mentions)
Total s6k, by Cell Signaling Technology (2 mentions)
Odyssey machine, by LI COR (2 mentions)
Ir 680 conjugated goat anti mouse or goat anti rabbit, by Thermo Fisher Scientific (2 mentions)
Irdye 800 conjugated donkey anti rabbit or donkey anti mouse, by LI COR (2 mentions)
γ tubulin, by Cell Signaling Technology (2 mentions)
P akt, by Cell Signaling Technology (2 mentions)
24 protocols using pparα
1
Antiobesity Effects of Aspergillus terreus
2
Immunochemical Analysis of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed, paraffin-embedded tissue sections were deparaffinized, rehydrated, and subjected to epitope retrieval using standard procedures. Antibodies against ColA1 (Santa Cruz Biotechnology), PGC-1α (Novus Biologicals), peroxisome proliferator-activated receptor alpha (PPAR-α; Cell Signaling Technology), and optic atrophy 1 (OPA1; Santa Cruz Biotechnology) were used for immunochemical staining. Sirius red staining was performed using the Sirius Red Staining Kit (Polysciences) according to the manufacturer’s protocol. Samples were examined under a laser-scanning microscope (Eclipse TE300, Nikon) [10 (link)].
3
Western Blot Analysis of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lysates from cells and tissues were subjected to 10 % SDS-PAGE (Bio-Rad, Pre-gel, MA); separated proteins were transferred onto Polyvinylidene difluoride (PVDF) membranes (Thermo Fisher Scientific, Seoul, Korea). Membranes were blocked with TBS-based blocking buffer (Li-COR Biosciences, Lincoln, NE) for 1h at room temperature, and incubated overnight at 4 °C with primary antibodies p16, SOD1,SOD2, ERK, p-ERK, CREB, p-CREB, p-elF2α, Beclin1, Bip, ATG12, LC3B, p62, BNIP3, PINK1, Parkin, PPARα, FATP4, Tau, P-tau, Beta-amyloid, GFAP, iba-1, beta-actin (1:1000, Cell Signaling Technology, Danvers, MA). Membranes were then washed with TBST three times for 10 min each and incubated with horseradish peroxidase-conjugated secondary antibodies (1:3000, Cell Signaling Technology, Danvers, MA) for 1 h at room temperature and washed again. Signals were detected by Odyssey-LC chemiluminsescent imaging system (LI-COR, Lincoln, NE). Signals were averaged and expressed as described in figure legend.
4
Western Blot Analysis of PPARα and A-FABP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells or quadriceps femoris muscles of mice were lysed with RIPA lysis buffer supplemented with 1% protease inhibitor cocktail. Equivalent amounts of supernatant samples were run on 10% acrylamide SDS-PAGE gel. Then the protein was transferred to nitrocellulose membrane (Millipore, Billerica, MA, USA). Membranes were blocked with 5% milk for 1 h and incubated with antibodies against PPARα (1:2000) (Cell Signaling Technology, Boston, MA), A-FABP (1:2000) (Cell Signaling Technology, Boston, MA) or β-actin (1:2000) (Cell Signaling Technology, Boston, MA) at 4°C overnight. Next, membranes were incubated with HRP-conjugated secondary antibody (1:3000) (Cell Signaling Technology, Boston, MA) for 1 h at room temperature. The protein bands were visualized with ECL detection kit (Cell Signaling Technology, Boston, MA). Densitometry analysis was performed using Image J software.
5
Western Blot Analysis of Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Sirt1, phospho-AMPK (Thr172), AMPK, FAS, SCD1, PPAR-α, and PPARδ, SREBP1, and TNF-α antibodies were obtained from Cell Signaling (Danvers, MA). Protein levels of cell extracts were measured by bicinchoninic acid assay (BCA) kit (Thermo Fisher Scientific Inc., Waltham, MA). For Western blot, 10–50 μg protein was resolved on 4–15% gradient polyacrylamide gels (criterion precast gel, Bio-Rad Laboratories, Hercules, CA), transferred to PVDF or nitrocellulose membranes, incubated in blocking buffer (5% nonfat dry milk in TBS), and then incubated with primary antibody (1 : 1000 dilution), washed, and incubated with horseradish peroxidase- or fluorescence-conjugated secondary antibody (1 : 10000 dilution). Visualization was conducted using Li-COR Odyssey Fc Imaging system (Li-COR Biosciences, Lincoln, NB) and band intensity was assessed using Quantity One (Bio-Rad Laboratories, Hercules, CA), with correction for background and loading controls.
6
Lipid Metabolism and Autophagy Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fetal bovine serum (FBS), culture media, TRIzol reagent, LysoTracker Red, BODIPY493/503, and the Lipofectamine 3000 transfection reagent were obtained from Invitrogen (Carlsbad, CA, USA). Palmitate (PA) and oleic acid (OA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against LC3, SQSTM1/p62, and TFE3 were obtained from Sigma-Aldrich. Antibodies against Cathepsin L, PGC1α, PPARα, CPT1α, and ACOX1 were from Cell Signaling Technology (Danvers, MA, USA). Antibodies against VPS11 and β-Actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Unless otherwise specified, all other reagents were purchased from Sigma-Aldrich.
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 were homogenized with 1× passive lysis buffer (Promega) supplemented with protease inhibitor cocktail (Sigma) and Xpert Phosphatase Inhibitor Cocktail Solution (GenDEPOT, Barker, TX, USA). Lysates were quantified using BCA protein assay kit (Thermo Fisher Scientific) and an equal amount of protein was electrophoresed on 8–10% SDS-polyacrylamide gel and transferred onto polyvinylidene fluoride membranes. Blots were blocked in 1× TBS-tween 20 containing 5% non-fat milk (Difco, Sparks, MD, USA) and incubated with primary antibodies. Antibodies including AMPK, p-AMPK, ACC, FAS, CPT1a, PEPCK-C, G6Pase, CD36, and PPAR-α were purchased from Cell signaling and α-tubulin, GAPDH were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Next day, the membranes were washed with 1× TBS-T and reacted with horseradish peroxidase-conjugated secondary antibodies (1:10,000) (Santa Cruz Biotechnology) and visualized by SuperSignalTM West Pico Chemiluminescent substrate (ThermoFisher Scientific). Band intensity was measured by Image J software 1.48v (NIH, USA) and was normalized to the α-tubulin.
8
Western Blot Analysis of Adipocyte Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were separated by 12% SDS–PAGE and transferred to nitrocellulose membranes and probed the membrane with primary antibodies against UCP1 (1:1,000), p‐PKA substrate (1:500), desnutrin (1:1,000), total Akt (1:1,000), phospho‐Akt (Ser473) (1:1,000), ERK1/2 (1:1,000), phosphorylated ERK1/2 (1:1,000), phosphorylated PKC α/β II (1:500) or HSL (1:1,000) (Cell Signaling), PPAR‐α (1:1,000), FADD (1:500), phosphorylated FADD (1:500), RIP140 (1:1,000) (Abcam), phosphorylated HSL (1:1,000) (Calbiochem), VLCAD (1:500), LCAD (1:1,000), MCAD (1:1,000), GAPDH (1:1,000), β‐actin (1:1,000), or α‐tubulin (1:1,000) (Santa Cruz Biotechnology).
9
Immunohistochemical Analysis of PPAR-α in Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lungs were fixed in 4% paraformaldehyde, embedded in paraffin and cut into sections of 4 μm in thickness. Sections were incubated with primary antibodies against PPAR-α (Cell Signaling Technology). After washing with phosphate-buffered saline (PBS), these sections were incubated with EnVision+HRP/Rb (DAKO, Glostrup, Denmark) at room temperature for 30 minutes. The positive signals of tissue sections were detected using the DAB (3,3′-diaminobenzidine) substrate kit (ZSGB-BIO, China) following the manufacturer’s instructions. The images were caught under a light microscope (Nikon, Tokyo, Japan).
10
Immunohistochemical Analysis of Lipid Metabolism Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tissue sections (5 μm) were subjected to antigen retrieval
by microwave after deparaffinization and rehydration for 10 min in
sodium citrate buffer. Sections were cooled to room temperature, treated
with 3% H2O2 for 10 min, and blocked with 5%
goat serum for 40 min at room temperature. One part of the sections
was stained with hematoxylin-eosin and oil red O ethanol dye, and
the other part was stained with immunohistochemistry. For immunohistochemistry
staining, the sections were incubated at 4 °C overnight with
the first antibody against PPAR-α, CPT1A, CPT2, SREBP-1C, and
CYP-7A (diluted 1:200, Cell Signaling Technology, Danvers, NA). Then,
the sections were washed in phosphate-buffered saline (PBS) and incubated
with the secondary antibody (biotinylated goat anti-rabbit, 1:150;
Vector Laboratories, Burlingame, CA) for 30 min. The sections were
counterstained with hematoxylin after diaminobenzidine staining. All
of the slices were dehydrated with ethanol for 2 min, followed by
xylene transparency for 5 min, and then the tablets were quickly sealed
with neutral gum and an ultrathin cover glass. After sealing the film,
it was observed under an ordinary optical microscope according to
our previous research methods.28 (link)
by microwave after deparaffinization and rehydration for 10 min in
sodium citrate buffer. Sections were cooled to room temperature, treated
with 3% H2O2 for 10 min, and blocked with 5%
goat serum for 40 min at room temperature. One part of the sections
was stained with hematoxylin-eosin and oil red O ethanol dye, and
the other part was stained with immunohistochemistry. For immunohistochemistry
staining, the sections were incubated at 4 °C overnight with
the first antibody against PPAR-α, CPT1A, CPT2, SREBP-1C, and
CYP-7A (diluted 1:200, Cell Signaling Technology, Danvers, NA). Then,
the sections were washed in phosphate-buffered saline (PBS) and incubated
with the secondary antibody (biotinylated goat anti-rabbit, 1:150;
Vector Laboratories, Burlingame, CA) for 30 min. The sections were
counterstained with hematoxylin after diaminobenzidine staining. All
of the slices were dehydrated with ethanol for 2 min, followed by
xylene transparency for 5 min, and then the tablets were quickly sealed
with neutral gum and an ultrathin cover glass. After sealing the film,
it was observed under an ordinary optical microscope according to
our previous research methods.28 (link)
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!