4 4 dianilino 1 1 binaphthyl 5 5 disulfonic acid dipotassium salt bis ans

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4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid, dipotassium salt (bis-ANS) is a fluorescent dye used in various laboratory applications. It functions as a probe to detect changes in protein structure and environments.

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5 protocols using 4 4 dianilino 1 1 binaphthyl 5 5 disulfonic acid dipotassium salt bis ans

Purification and Characterization of Human αA and αB Crystallins

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Insulin, citrate synthase, protease inhibitor cocktail, dithiothreitol, trinitrobenzenesulfonic acid (TNBS), 4,4′-dianilino-1,1′-binaphthyl-5,5′-disulfonic acid (bis-ANS), dipotassium salt, and succinyl coenzyme A (SuccCoA) were obtained from Sigma-Aldrich (St. Louis, MO). N^ε-Acetyllysine was obtained from Chem-Impex International Inc. (Wood Dale, IL). All other chemicals were of analytical grade.
Cloning, expression, and purification of human αA and αB were performed as previously described.^{24 (link),25}

Nandi S.K., Rakete S., Nahomi R.B., Michel C., Dunbar A., Fritz K.S, & Nagaraj R.H. (2019). Succinylation Is a Gain-of-Function Modification in Human Lens αB-Crystallin. Biochemistry, 58(9), 1260-1274.

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Amyloid Aggregation Assay Protocol

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4,4′-Dianilino-1,1′-binaphthyl-5,5′-disulfonic acid (bisANS) dipotassium salt, alpha-lactalbumin, bovine serum albumin (BSA), citrate synthase, 2-mercaptoethanol, sodium butyrate, thioflavin T (ThioT) and Greiner 384-well clear flat bottom plates were purchased from Sigma Aldrich (Australia). Pluronic F-68 (PF-68) was purchased from Thermo Fisher Scientific (Australia). Human beta-amyloid (1–42) was purchased from AnaSpec (Australia). Tryptone N1 was a kind gift from Organotechnie (France). All other chemicals were of analytical grade and purchased from Sigma Aldrich (Australia) unless otherwise specified.

Satapathy S., Dabbs R.A, & Wilson M.R. (2020). Rapid high-yield expression and purification of fully post-translationally modified recombinant clusterin and mutants. Scientific Reports, 10, 14243.

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Fluorescence-based Enzyme Kinetics

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Bovine insulin, citrate synthase (CS) from porcine heart, Dithiothreitol (DTT) and 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid, dipotassium salt (bis-ANS) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Alexa fluor 350 and Alexa fluor 488 protein labeling kits (Catalog Nos. A-10170 and A-10235, respectively) were obtained from Molecular Probes (Invitrogen, Carlsbad, CA, USA). Malate dehydrogenase (MDH) from porcine heart, disodium hydrogen phosphate and mono sodium hydrogen phosphate were from Sisco Research Laboratories, India. All other chemicals were of analytical grade.

Nandi S.K., Panda A.K., Chakraborty A., Ray S.S, & Biswas A. (2015). Role of Subunit Exchange and Electrostatic Interactions on the Chaperone Activity of Mycobacterium leprae HSP18. PLoS ONE, 10(6), e0129734.

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Biochemical Assays for Enzyme Kinetics

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Dithiothreitol (DTT), 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid dipotassium salt (bis-ANS), urea, isopropyl β-D-1-thiogalactopyranoside (IPTG), adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine 5′-[γ-thio] triphosphate tetralithium salt (ATP-γS), 2′,3′-O-(2,4,6-Trinitrophenyl) adenosine 5′-triphosphate tetrasodium salt (TNP-ATP), lysozyme, citrate synthase (CS), were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Malate dehydrogenase (MDH), oxaloacetic acid (OAA), nicotinamide adenine dinucleotide (reduced) disodium salt (NADH), trypsin, buffer salts (phosphate, tris, etc.) were from Sisco Research Laboratories, India. All other chemicals were of analytical grade.

Nandi S.K., Chakraborty A., Panda A.K., Sinha Ray S., Kar R.K., Bhunia A, & Biswas A. (2015). Interaction of ATP with a Small Heat Shock Protein from Mycobacterium leprae: Effect on Its Structure and Function. PLoS Neglected Tropical Diseases, 9(3), e0003661.

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ApoA-I Recombinant Protein Expression

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Escherichia coli strains used in this study were maintained and stored using the routine culture techniques. The pMD18-T vector containing the full-length apoA-I cDNA was purchased from Sino Biological Inc. (China). The pET28a (+) vector was from Novagen (Madison, WI). The culture media components were purchased from HiMedia (Mumbai, India). The enzymes and chemicals used for DNA manipulation were obtained from Thermo Scientific (USA). The 6 × His-purifying resin was from Novagen. The oligonuleotide primers, guanidine hydrochloride (GndHCl), thioflavinT (ThT), 4,4′-dianilino-1,1′-binaphthyl-5,5′-disulfonic acid dipotassium salt (bis-ANS), congo red, sodium dodecyl sulphate (SDS), agarose, anti-His tag (catalog no. H1029) and HRP-conjugated anti-mouse IgG were procured from Sigma-Aldrich (St Louis, MO). 1,2-dimyristoyl sn-glycero-3-phosphatidylcholine (DMPC) was from Avanti Polar Lipids (Alabaster, AL). The QuickChange II XL Site-Directed Muatgenesis kit was from Stratagene (LaJolla, CA). Coomassie Brilliant Blue R250 (CBB-R250) was obtained from Merck. Other reagents were of the highest analytical grade available, unless otherwise stated.

Ma'arfi F., Chandra S., Fatima J.E., Khan M.Y., Mir S.S, & Yusuf M.A. (2020). Probing the Structure–Function relationship and amyloidogenic propensities in natural variants of apolipoprotein A-I. Biochemistry and Biophysics Reports, 24, 100815.

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