All reagents were used without further treatment. Fmoc-protected amino acids were purchased from GL Biochem. 2-(6-Chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate (HCTU), trifluoroacetic acid (TFA), and hydroxybenzotriazole hydrate (HOBt) were purchased from Chem-Impex International. 4-methylpiperidine were purchased from Acros Organics. Rink Amide-ChemMatrix resin (0.5 mmol/g loading) was purchased from Biotage. All other reagents including tris(2-carboxyethyl)phosphine hydrochloride (TCEP), DL-Dithiothreitol (DTT), N,N-Dimethyldodecylamine N-oxide (LDAO), N-Dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (C12 Betaine), Triton X100 were purchased from Sigma-Aldrich. Fos-choline-12 (DPC), Decyl β-D-maltopyranoside (DM) were purchased from Anatrace. Sodium dodecyl sulfate (SDS) was purchased from Calbiochem, IPTG (isopropyl β-D-1-thiogalactopyranoside) was purchased from Genesee Scientific Corporation.
N n dimethyldodecylamine n oxide ldao
Manufactured by Merck Group
N,N-Dimethyldodecylamine N-oxide (LDAO) is a chemical compound commonly used as a surfactant in laboratory applications. It functions as a mild, non-ionic detergent with the ability to solubilize and maintain the stability of various biomolecules, such as proteins and membranes, in aqueous solutions.
Automatically generated - may contain errors
Lab products found in correlation
4 methylpiperidine, by Thermo Fisher Scientific (2 mentions)
Fos choline 12, by Anatrace (2 mentions)
Decyl β d maltopyranoside, by Anatrace (2 mentions)
2 6 chloro 1h benzotriazole 1 yl 1 1 3 3 tetramethylaminium hexafluorophosphate hctu trifluoroacetic acid tfa, by Chem-Impex (2 mentions)
Hydroxybenzotriazole hydrate hobt, by Chem-Impex (2 mentions)
N dodecyl n n dimethyl 3 ammonio 1 propanesulfonate c12 betaine triton x100, by Merck Group (2 mentions)
Fmoc protected amino acids, by GL Biochem (2 mentions)
Rink amide chemmatrix resin, by Biotage (2 mentions)
N dodecyl β d maltoside, by Anatrace (2 mentions)
Ppicza, by Thermo Fisher Scientific (1 mentions)
Cm5 sensor chip, by GE Healthcare (1 mentions)
Zero blunt topo pcr cloning kit, by Thermo Fisher Scientific (1 mentions)
Biacore 3000, by GE Healthcare (1 mentions)
N decyl β d maltoside, by Anatrace (1 mentions)
Zeocin, by InvivoGen (1 mentions)
Cholesteryl hemisuccinate, by Merck Group (1 mentions)
Ethanolamine, by GE Healthcare (1 mentions)
Hrv 3c protease, by Sino Biological (1 mentions)
Fos choline 12 fc12, by Anatrace (1 mentions)
Triton x 100, by Merck Group (1 mentions)
5 protocols using n n dimethyldodecylamine n oxide ldao
1
Peptide Synthesis and Purification Protocol
2
Heterologous Expression of Human STRA6
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Pichia pastoris strain KM71H (aox::ARG4, arg4), expression vector pPICZ-A and Zero Blunt TOPO PCR cloning kit were purchased from Invitrogen. A plasmid containing the gene for full-length human STRA6 transcript variant 2 (NM_022369.3) in the pCMV6-XL4 cloning vector, was purchased from OriGene. The pEGFP-N1 plasmid containing eGFP was purchased from Clontech. Zeocin was from Invivogen. DyLight 594 amine-reactive dye was purchased from Pierce. N-dodecyl β-D maltoside (DDM), N-decyl β-D maltoside (DM) and Fos-Choline-12 (FC-12) were from Anatrace. Cholesteryl hemisuccinate, N, N-dimethyldodecylamine N-oxide (LDAO) and dodecyl nonaoxyethylene ether (C12E9) were from Sigma. HRV 3C protease was from Sino Biological. Polyclonal antibody raised against full-length human STRA6 (#H00064220-D01P) was from Abnova. The Biacore 3000 and instrument-specific consumables (CM5 sensor chip, ethyl(dimethylaminopropyl)-carbodiimide, N-hydroxysulphosuccinimide and ethanolamine) were obtained from GE Healthcare.
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Pichia pastoris strain KM71H (aox::ARG4, arg4), expression vector pPICZ-A and Zero Blunt TOPO PCR cloning kit were purchased from Invitrogen. A plasmid containing the gene for full-length human STRA6 transcript variant 2 (NM_022369.3) in the pCMV6-XL4 cloning vector, was purchased from OriGene. The pEGFP-N1 plasmid containing eGFP was purchased from Clontech. Zeocin was from Invivogen. DyLight 594 amine-reactive dye was purchased from Pierce. N-dodecyl β-D maltoside (DDM), N-decyl β-D maltoside (DM) and Fos-Choline-12 (FC-12) were from Anatrace. Cholesteryl hemisuccinate, N, N-dimethyldodecylamine N-oxide (LDAO) and dodecyl nonaoxyethylene ether (C12E9) were from Sigma. HRV 3C protease was from Sino Biological. Polyclonal antibody raised against full-length human STRA6 (#H00064220-D01P) was from Abnova. The Biacore 3000 and instrument-specific consumables (CM5 sensor chip, ethyl(dimethylaminopropyl)-carbodiimide, N-hydroxysulphosuccinimide and ethanolamine) were obtained from GE Healthcare.
3
Peptidisc Purification and Characterization
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Peptidisc peptides were obtained from Peptidisc Biotech. Tryptone, yeast extract, Tris-base, imidazole, NaCl, dithiothreitol (DTT), acrylamide 40%, bis-acrylamide 2% and TEMED were obtained from Bioshop, Canada. Isopropyl β-D-1-thiogalactopyranoside (IPTG), ampicillin, kanamycin and arabinose were purchased from GoldBio, EDTA-free protease inhibitor cocktail from Roche, n-dodecyl-β-D-maltoside (DDM) and octyl-β-D-glucoside (β-OG) from Anatrace, and N,N-dimethyldodecylamine N-oxide (LDAO) and Triton X-100 from Sigma. Superdex 200 HR and Superose 6 resins were obtained from GE Healthcare. Most other chemicals were purchased from Fisher Scientific.
4
Circular Dichroism Spectroscopy of Proteins
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CD spectra were recorded on an Aviv Model 410 (Aviv Biomedical Inc., Lakewood, NJ) at 25 °C using a 0.1-cm quartz cuvette. Protein was prepared in either 10 mm potassium phosphate buffer, pH 7.4, or 25 mm Tris, pH 7.4, 1 mm MgCl2 with or without 0.05% (v/v) N,N-dimethyldodecylamine N-oxide (LDAO; Sigma) at concentrations ranging from 2.6 to 10 μm . Spectra were recorded between 190 and 260 nm in increments of 1 nm, with a bandwidth of 1 nm and an averaging time of 5 s. Spectra from three scans were averaged, baseline corrected, and smoothed using the Aviv software. Spectra are expressed as mean residue ellipticity using protein concentrations determined by amino acid analysis. CD spectra were deconvoluted with the CDSSTR algorithm using the Dichroweb software (58 (link), 59 (link)). The estimated helical content given was obtained when using protein reference set 4 (60 (link)). Normalized mean root square deviation was <0.011 for all deconvolutions described.
5
Peptide Synthesis and Purification Protocol
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