Atpenin a5

Manufactured by Santa Cruz Biotechnology

Atpenin A5 is a natural product isolated from the fungus Penicillium sp. that inhibits the mitochondrial respiratory chain complex II (succinate dehydrogenase). It is a useful tool for the study of cellular metabolism and the role of mitochondrial respiration in various biological processes.

Automatically generated - may contain errors

Lab products found in correlation

7 protocols using atpenin a5

Mitochondrial Dynamics Protein Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies to the following proteins were used: Drp1 (BD Biosciences), Drp1 S637-PO₄ (Cell Signaling), Actin (Millipore), Fis1 (Alexis Corps), Mff (Sigma-Aldrich), Mfn1 (Chen et al., 2003 (link)), Mfn2 (Sigma), Opa1 (in house monoclonal), ClpP (Proteintech Group), Oxa1 (Proteintech Group), Tom20 (Santa Cruz BioTech), Hsp60 (Santa Cruz BioTech), Yme1L (Proteintech Group), Myc (Covance). Oligomycin (Sigma-Aldrich), antimycin A (Sigma-Aldrich), atpenin A5 (Santa Cruz BioTech), CCCP (Sigma-Aldrich) and atractyloside (Sigma-Aldrich) were used at 1 μM. Concentrations of other compounds: rotenone (Sigma-Aldrich), 200 nM; KCN (Sigma-Aldrich), 1 mM; o-phenanthroline (Sigma-Aldrich), 0.5 mM; aspartate, 1 mM; ADP (Sigma-Aldrich), 200 μM; β-NADH (Sigma-Aldrich), 2 mM. Unless otherwise indicated, substrates (pyruvate, malate, succinate, α-ketoglutarate) were used at 5 mM.

Mishra P., Carelli V., Manfredi G, & Chan D.C. (2014). Proteolytic cleavage of Opa1 stimulates mitochondrial inner membrane fusion and couples fusion to oxidative phosphorylation. Cell metabolism, 19(4), 630-641.

+ Open protocol

+ Expand

Screening for Mitochondrial Modulators

Check if the same lab product or an alternative is used in the 5 most similar protocols

CaCl₂ standard was from Thermo Scientific, fatty acid-free BSA from Calbiochem, and atpenin A5 from Santa Cruz Biotechnology. Amplex UltraRed, tetramethylrhodamine methyl ester (TMRM), LysoTracker Red DND-99, and Geltrex were from Invitrogen. Compounds for screening were arbitrarily selected from a diverse, non-combinatorial library obtained from ChemBridge (∼10 mM in DMSO). New stocks of primary hits and their structural analogs were also sourced from ChemBridge (and dissolved to 80 mM in DMSO). The biological activity of iGP-1 (ChemBridge ID 5224148) was validated using compound obtained from Vitas-M (ID STK017597) and found to be essentially identical. All other reagents were from Sigma-Aldrich. sn-Glycerol 3-phosphate was added as disodium rac-α/β-glycerol phosphate (25% active optical isomer sn-glycerol 3-phosphate, 25% inactive optical isomer sn-glycerol 1-phosphate and 50% inactive structural isomer glycerol 2-phosphate) and will be referred to as glycerol phosphate. Where indicated, total and free calcium concentrations in assay buffers were calculated using the Extended MaxChelator program available through Chris Patton on the Stanford University Web site at http://maxchelator.stanford.edu.

Orr A.L., Ashok D., Sarantos M.R., Ng R., Shi T., Gerencser A.A., Hughes R.E, & Brand M.D. (2014). Novel Inhibitors of Mitochondrial sn-Glycerol 3-phosphate Dehydrogenase. PLoS ONE, 9(2), e89938.

+ Open protocol

+ Expand

Mitochondrial Respiratory Chain Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rotenone, malonate, antimycin A, NaN₃, oligomycin, glutamate, malate, succinate, ascorbate, N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD), ADP, ATP-Mg, adenylyl-imidodiphosphate (AMP-PNP), pericidin A, and myxothiazol were from Sigma-Aldrich. Atpenin A5 was from Santa Cruz Biotechnology. Potassium cyanide (KCN) was from the Department of Chemistry, Peking University. Tetramethylrhodamine methyl ester (TMRM) was from Molecular Probes (Thermo Fisher Scientific). Other reagents were obtained commercially at the highest grade available.

Feng G., Liu B., Li J., Cheng T., Huang Z., Wang X, & Cheng H.(. (2019). Mitoflash biogenesis and its role in the autoregulation of mitochondrial proton electrochemical potential. The Journal of General Physiology, 151(6), 727-737.

+ Open protocol

+ Expand

Mitochondrial Electron Transport Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rotenone, 2-thenoyltrifluoroacetone, antimycin A, myxothiazol, potassium cyanide, 3,4-dehydro-l-proline, carboxin, methyl-succinate, l-proline, coenzyme Q1, doxycycline, N-acetyl cysteine, and o-aminobenzaldehyde were purchased from Sigma Aldrich, St. Louis, MO. Atpenin A5 was purchased from Santa Cruz Biotechnology, Dallas, Texas.

Hancock C.N., Liu W., Alvord W.G, & Phang J.M. (2015). Co-regulation of mitochondrial respiration by proline dehydrogenase/oxidase and succinate. Amino Acids, 48, 859-872.

+ Open protocol

+ Expand

Synthesis and Characterization of GB111-NH2 Analogs

Check if the same lab product or an alternative is used in the 5 most similar protocols

See below for synthesis and characterization of GB111-NH₂ and analogs. NMR spectra were recorded on a Varian 400 MHz (400/100) or a Varian Inova 500 MHz (500/126 MHz) equipped with a pulsed field gradient accessory. Chemical shifts (∂) are reported in parts per million (ppm) downfield from tetramethylsilane and are reference to the residual protium signal in the NMR solvents. Data are reported as follows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet, m=multiplet and q=quartet), coupling constant (J) in Hertz (Hz) and integration. E64d (Enzo Lifesciences, Farmingdale NY), leupeptin (Sigma Aldrich, St. Louis MO), CA074Me (EMD Millipore, Hayward CA), LPS from E. coli 0111:B4 (Sigma Aldrich), 6-aminonicotinamide (Santa Cruz Biotech, Santa Cruz CA), Imject Alum (Pierce Biotechnology, Rockford IL), ATPenin A5 (Santa Cruz Biotech), N-ethylmaleimide (Sigma), rotenone (Sigma), oligomycin A (Cayman Chemical, Ann Arbor MI), nigericin (Cayman Chemical), MitoSOX (Life Technologies, Carlsbad CA), ATP (Sigma), and koningic acid (Adipogen, Switzerland) were purchased from commercial sources, dissolved in vendor-recommended solvents, and used without further purification. ENOblock (Jung et al., 2013 (link)) was a generous gift from Dr. Darren Williams.

Sanman L.E., Qian Y., Eisele N.A., Ng T.M., van der Linden W.A., Monack D.M., Weerapana E, & Bogyo M. (2016). Disruption of glycolytic flux is a signal for inflammasome signaling and pyroptotic cell death. eLife, 5, e13663.

+ Open protocol

+ Expand

Chemical Modulators for Cellular Metabolism

Check if the same lab product or an alternative is used in the 5 most similar protocols

The heme inhibitor succinylacetone (SA) (Sigma, D1415) is used at a concentration of 0.5 mM. Hemin (Sigma, 51280) is used at a concentration of 10 μM in 0.1 N NaOH. Diethyl butylmalonate (BM) (Sigma, 112038) is used at a concentration of 1 mM. Atpenin A5 (Santa Cruz biotechnology, sc-202475) is used at 250 nM. SIS3 (Selleck chemicals, S7959) is used at 5 μM. NF-56-EJ40 (Axon Medchem 3056) is used at 1 μM. AZD3965 (Selleckchem S7339) is used at 1 μM. Exogenous succinate is provided as either 40 mM Sodium succinate (Sigma) or 16 mM dimethylsuccinate (Sigma) supplementation. HDM inhibition was achieved with 250 nM JIB04 (Medchem express HY-13953) and TET inhibition with 5 μM TETin-C35 (Aobious AOB11121). For retinoic acid experiments, RA, HY-14649, HX531 (HY-108521) and AGN193109 (HY-U00449) were used at 0.5 µM, 10 µM and 100 nM, respectively (all from Medchemexpress). Inhibition of Sirt7 is achieved using 5 µM of the Sirt7 inhibitor 97491 (MedChemExpress).

Detraux D., Caruso M., Feller L., Fransolet M., Meurant S., Mathieu J., Arnould T, & Renard P. (2023). A critical role for heme synthesis and succinate in the regulation of pluripotent states transitions. eLife, 12, e78546.

+ Open protocol

+ Expand

Mitochondrial Protein and Function Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Acrylamide solution, Laemmli buffer, tetramethylethylenediamine, and nitrocellulose membrane were obtained from Bio-Rad. Goat anti-mouse antibody, oxidative phosphorylation (OXPHOS) primary antibody cocktail, PDH primary antibody cocktail, and UCP3 primary antibody were obtained from Abcam. The triglyceride assay kit and insulin ELISA kit were obtained from Cederlane and Millipore, respectively. Amplex UltraRed (AUR) reagent and 100 bp protein ladder were obtained from Invitrogen. Atpenin A5, goat anti-rabbit antibody, NDUSF1 primary antibody, and superoxide dismutase (SOD)2 primary antibody were obtained from Santa Cruz. Adenosine diphosphate (ADP), antimycin A, bovine serum albumin (BSA), disulfiram, ethylene glycolbis(b-aminoethyl ether)-N,N,N¢,N¢-tetraacetic acid (EGTA), GDP, genapin, glutathione assay kit, glycine, HEPES, horseradish peroxidase (HRP), KMV, malic acid, mannitol, MgCl 2 , NaCl, NADH, oligomycin, palmitoyl carnitine, ponceau, pyruvic acid, the red Extract-N-Amp PCR kit, rotenone, SOD, sodium azide, subtilisin A, succinic acid, and sucrose were all obtained from Sigma.

Young A., Gardiner D., Kuksal N., Gill R., O'Brien M, & Mailloux R.J. (2019). Deletion of the Glutaredoxin-2 Gene Protects Mice from Diet-Induced Weight Gain, Which Correlates with Increased Mitochondrial Respiration and Proton Leaks in Skeletal Muscle. Antioxidants & redox signaling, 31(17).

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!