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Alamethicin

Manufactured by Cayman Chemical
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Alamethicin is a fungal-derived peptide that exhibits ion channel-forming activity. It has been utilized as a laboratory tool to study ion transport and membrane permeabilization processes.

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Lab products found in correlation

Pageblue protein staining solution, by Thermo Fisher Scientific (2 mentions) Atpenin a5, by Cayman Chemical (2 mentions) Nativemark unstained protein standard, by Thermo Fisher Scientific (2 mentions) Amplex ultrared, by Thermo Fisher Scientific (2 mentions) Triphenyltetrazolium chloride, by Merck Group (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Piericidin a, by Merck Group (1 mentions) Benserazide, by Merck Group (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Catalase, by Merck Group (1 mentions) Menadione, by Merck Group (1 mentions) Ros glo h2o2 assay, by Promega (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Pyrogallol, by Merck Group (1 mentions) Celltiter glo, by Promega (1 mentions) Horseradish peroxidase, by Thermo Fisher Scientific (1 mentions) Mannitol, by Merck Group (1 mentions) Hexaammineruthenium 3 chloride, by Merck Group (1 mentions)

5 protocols using alamethicin

1

Flavin-Site Assay for R-CI Orientation

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R-CI content and orientation in PLs were determined using the NADH:APAD+ oxidoreduction assay, which reports on the activity of the flavin (NADH oxidation) site of complex I in the presence of 100 μM NADH (Alfa Aesar), 500 μM APAD+ (Sigma-Aldrich) and 1 μM piericidin A (Sigma-Aldrich).14 (link) Note that lipid composition does not affect flavin-site activity,41,42 (link) but rather the efficiency of incorporating R-CI into liposomes. To assess the orientation of R-CI, the assay was conducted in the presence/absence of the pore-forming antibiotic alamethicin (20 μg mL−1, Cayman Chemical). Absorbance was measured at 400 nm and 450 nm (ε400–450 = 3.16 mM−1 cm−1).
Eisermann J., Wright J.J., Wilton-Ely J.D., Hirst J, & Roessler M.M. (2023). Using light scattering to assess how phospholipid–protein interactions affect complex I functionality in liposomes. RSC Chemical Biology, 4(6), 386-398.
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2

Oxidative Stress and Cell Viability Assays

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All materials were reagent grade. Minimal essential media (MEM) Glutamax, 100X antibiotic/antimycotic, heat-inactivated FBS, 100 mM sodium pyruvate, and 0.25% Trypsin–EDTA were from Life Technologies (Grand Island, NY, USA). Alamethicin was from Cayman Chemical (Ann Arbor, MI, USA). DMSO, menadione, eseroline (fumarate salt), benserazide, pyrogallol, and catalase were from Sigma-Aldrich (St. Louis, MO, USA). CellTiter-Glo® and ROS-Glo™ H2O2 Assays were from Promega Corp. (Madison, WI, USA).
Kelts J.L., Cali J.J., Duellman S.J, & Shultz J. (2015). Altered cytotoxicity of ROS-inducing compounds by sodium pyruvate in cell culture medium depends on the location of ROS generation. SpringerPlus, 4, 269.
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3

Biochemical Assays for Oxidoreductase Activity

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Chemicals were purchased from Sigma: bovine serum albumin (BSA) essentially fatty acid free (#A6003), DDM (#D4641), hexaammineruthenium (III) chloride (#262005), horseradish peroxidase (#P8250), mannitol (#63559), NADH (#N8129), Q1 (#C7956), SOD (#S9697), sucrose (#S7903), and triphenyl-tetrazolium chloride (#T8877). 3–12% acrylamide gradient gel (#BN1001BOX), Amplex UltraRed (#A36006), NativeMark™ unstained protein standards (#LC0725), PageBlue Protein Staining Solution (#24620), and Pierce BCA protein assay kit (#23225) were from Thermo Fisher Scientific. Alamethicin (#11425) and atpenin A5 (#11898) were from Cayman Chemical.
Yoval-Sánchez B., Ansari F., James J., Niatsetskaya Z., Sosunov S., Filipenko P., Tikhonova I.G., Ten V., Wittig I., Rafikov R, & Galkin A. (2022). Redox-dependent loss of flavin by mitochondria complex I is different in brain and heart. Redox Biology, 51, 102258.
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4

Membrane Protein Purification and Characterization

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Most of the chemicals were purchased from Sigma, including mannitol (catalog no. 63559), sucrose (catalog no. 84097), essentially fatty acid–free bovine serum albumin (BSA; catalog no. A6003), NADH (catalog no. N8129), hexaammineruthenium (III) chloride (catalog no. 262005), Q1 (catalog no. C7956), DDM (catalog no. S#D4641), and triphenyl-tetrazolium chloride (catalog no. T8877). Pierce bicinchoninic acid protein assay kit (catalog no. 23225), Amplex UltraRed (catalog no. A36006), horseradish peroxidase (catalog no. 012001), 3 to 12% acrylamide gradient gel, 8 × 8 cm (catalog no. BN1001BOX), NativeMark unstained protein standards (catalog no. LC0725), and PageBlue Protein Staining Solution (catalog no. 24620) were from Thermo Fisher Scientific. Alamethicin (catalog no. 11425) and atpenin A5 (catalog no. 11898) were from Cayman Chemical.
Ansari F., Yoval-Sánchez B., Niatsetskaya Z., Sosunov S., Stepanova A., Garcia C., Owusu-Ansah E., Ten V., Wittig I, & Galkin A. (2021). Quantification of NADH:ubiquinone oxidoreductase (complex I) content in biological samples. The Journal of Biological Chemistry, 297(4), 101204.
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5

Enzymatic Assay for Glycerol-1-Phosphate

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The following chemicals were purchased from Sigma: adenosine 5′-phosphate monopotassium salt hydrate (#A5285), alcohol dehydrogenase (#A7011), aminocaproic acid (#A2504), DDM (#D4641), EDTA (#E–5391), FMN (#F2253), imidazole (#792527), L-Malic acid disodium salt monohydrate (#233935), NADH (#N8129), phenylmethylsulfonyl fluoride (PMSF) (#P7626), rac-glycerol-1-phosphate sodium salt (#61668), SDS (#L3771), sodium pyruvate (#P5280), sodium succinate dibasic (#14160), and triphenyl-tetrazolium chloride (#T8877). NativeMark unstained protein standards (#LC0725), Pierce BCA protein assay kit (#23225), and 3–12% acrylamide gradient gel, 8 × 8 cm (#BN1001BOX) were from Thermo Fisher Scientific. Alamethicin (#11425), metformin (#13118), and N-ethylmaleimide (NEM) (#19938) were from Cayman Chemical. Difluorocarboxyfluorescein NHS-ester was from Fluoroprobes (#1223).
Yoval-Sánchez B., Ansari F., Lange D, & Galkin A. (2022). Effect of metformin on intact mitochondria from liver and brain: Concept revisited. European journal of pharmacology, 931, 175177.
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