Bicinchoninic acid assay bca protein assay kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Bicinchoninic acid assay (BCA) protein assay kit is a colorimetric detection and quantification method for measuring the total protein concentration in a sample. The assay is based on the reduction of Cu2+ to Cu+ by protein in an alkaline medium, and the subsequent chelation of the Cu+ ion with BCA, producing a purple-colored complex that absorbs light at 562 nm.

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6 protocols using bicinchoninic acid assay bca protein assay kit

Multiplex eSimoa Immunoassay Protocol

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All antibodies and recombinant proteins used in the eSimoa assays are listed in Table S1 (Supporting Information). All the buffers, dye‐encoded carboxylated paramagnetic beads, and consumables used in the eSimoa assays were purchased from Quanterix Corporation. Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), antibiotics (10000 U mL⁻¹ penicillin, 10000 µg mL⁻¹ streptomycin, and 29.2 mg mL⁻¹ L‐glutamine), Dulbecco's phosphate‐buffered saline (DPBS), phosphate‐buffered saline (PBS) and trypsin‐ethylenediaminetetraacetic acid (trypsin‐EDTA) (0.05%) were purchased from Gibco. Tween‐20, sodium chloride, ethylenediaminetetraacetic acid (EDTA), sodium deoxycholate, and sodium dodecyl sulfate were purchased from Sigma‐Aldrich. Bicinchoninic acid (BCA) assay protein assay kit and phenylmethylsulfonyl fluoride (PMSF) were purchased from Thermo Fisher Scientific. Tris hydrochloride (Tris HCl) was purchased from iNtRON Biotechnology. Laemmli sample buffer and Clarity Western ECL Substrate were purchased from Bio‐Rad. The antibodies used for western blotting analysis were listed in Characterization. Proteinase K was purchased from Invitrogen. Streptavidin‐coated silica bead was purchased from Bangs Laboratories. Sepharose CL‐6B resin was purchased from GE Healthcare/Cytiva.

Cheng C., Hou K., Hsu C, & Chiang L. (2023). Ultrasensitive and High‐Resolution Protein Spatially Decoding Framework for Tumor Extracellular Vesicles. Advanced Science, 11(3), 2304926.

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Liposome-based Blood-Brain Barrier Delivery

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Losartan potassium was obtained from the Tokyo chemical industry (Tokyo, Japan), Dioleoyl-3-trimethylammonium-propane chloride (DOTAP), Lissamine rhodamine and dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were bought from Avanti Polar Lipids (Birmingham, AL, USA). DSPE–PEG₂₀₀₀–NHS was purchased from Biochempeg Scientific Inc. (Watertown, MA, USA). Bicinchoninic acid (BCA) assay protein assay kit was bought from Thermo Fisher Scientific. (Waltham, MA, USA). Holo-transferrin, Cholesterol, Hoechst 33342, ethylenediaminetetraacetic acid (EDTA), 4-(2-hydroxyethyl)-1- piperazineethanesulfonic acid (HEPES), hCMEC/D3 complete Media and Triton X-100 were procured from Sigma-Aldrich (St. Louis, MO, USA). Phosphate buffered saline (PBS), fetal bovine serum (FBS), and Dulbecco’s modified Eagle medium (DMEM) were acquired from Corning Incorporated (Corning, NY, USA). Penetratin (RQIKIWFQNRRMKWKKGG) was purchased from Zhejiang Ontores Biotechnologies Co., Ltd. (Zhejiang, China).

Optiz T., Grooms S.Y., Bennett M.V, & Zukin R.S. (2000). Remodeling of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor subunit composition in hippocampal neurons after global ischemia. Proceedings of the National Academy of Sciences of the United States of America, 97(24), 13360-13365.

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Polymer-Based Insulin Delivery System for Diabetes

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PVA (Mw = 31,000–50,000 Da; 87–89% hydrolysed), PEG flakes (Mw = 2000 Da), sodium tetrahydroxyborate decahydrate, insulin (recombinant human; dry powder), poly(D,L-lactide-co-glycolide) (acid terminated; lactide/glycolide 50:50; Mw = 24,000–38,000) and streptozotocin (STZ) were also purchased from Sigma-Aldrich Ltd., Poole, UK. Dulbecco’s phosphate-buffered saline (DPBS) and sodium hydroxide (NaOH) were purchased from Fisher Scientific, Loughborough, UK. A bicinchoninic acid assay (BCA) protein assay kit was purchased from Thermo Fisher Scientific, Pierce Biotechnology Inc., Rockford, USA. Trifluoroacetic acid (TFA), hydrochloric acid (HCl) solution, acetonitrile, dichloromethane (DCM) and methanol were of HPLC grade. All other reagents and solvents were of appropriate laboratory standard and used without further purification.

Abdelkader D.H., Tambuwala M.M., Mitchell C.A., Osman M.A., El-Gizawy S.A., Faheem A.M., El-Tanani M, & McCarron P.A. (2018). Enhanced cutaneous wound healing in rats following topical delivery of insulin-loaded nanoparticles embedded in poly(vinyl alcohol)-borate hydrogels. Drug Delivery and Translational Research, 8(5), 1053-1065.

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Lipid Nanoparticle-Based Gene Delivery

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In vivo-jetPEI (linear 22-kDa PEI; in vivo-jetPEI polyethylenimine [lPEI]) was purchased from Polyplus Transfection (Illkirch, France). Branched PEI with molecular weight 10,000 Da (bPEI10k) was bought from Alfa Aesar (Wardhill, MA, USA). 1,2-Di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and cholesterol were purchased from Avanti Polar Lipids (Alabaster, AL, USA). 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG2000) was purchased from NOF America Corporation (White Plains, NY, USA). LF3000, TRIzol RNA isolation reagent, and bicinchoninic acid assay (BCA) protein assay kit were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Direct-zol RNA MiniPrep Kit was from Zymo Research (Irvine, CA, USA). Celltiter-Glo assay was purchased from Promega (Madison, WI, USA). Matrigel was purchased from Corning (Corning, NY, USA). Human AFP ELISA kit was purchased from R&D Systems (San Diego, CA, USA). All other chemicals and organic solvents of analytical grade were purchased from Sigma-Aldrich or Thermo Fisher Scientific.

Jilek J.L., Zhang Q.Y., Tu M.J., Ho P.Y., Duan Z., Qiu J.X, & Yu A.M. (2019). Bioengineered Let-7c Inhibits Orthotopic Hepatocellular Carcinoma and Improves Overall Survival with Minimal Immunogenicity. Molecular Therapy. Nucleic Acids, 14, 498-508.

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Luciferase Activity Quantification

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Twenty-four hours post transfection, the culture medium was removed, cells were gently washed with PBS once, and 100 μl of passive lysis buffer (PLB) was added and incubated at 37 °C for 5 min. Cells were lysed by pipetting up and down several times and spun at 5000 rpm for 4 min to remove debris, and 10–20 μl was used to assay for luciferase activity using luciferase assay system (Promega) in a single injector luminometer. Total protein concentration was measured using bicinchoninic acid assay (BCA) protein assay kit (Thermo Fisher), and relative light units were normalized to micrograms of protein. Graphs were plotted from data obtained as a mean of three independent experiments with standard deviation as error bars.

Lutgen V., Narasipura S.D., Sharma A., Min S, & Al-Harthi L. (2016). β-Catenin signaling positively regulates glutamate uptake and metabolism in astrocytes. Journal of Neuroinflammation, 13(1), 242.

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Kunming Mice Protein Isolation Protocol

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Male Kunming mice weighing 25-30 g were purchased from the Laboratory Animal Center of Southern Medical University (Guangzhou, China). The animal experiments were approved by the local Animal Ethics Committee of the Southern Medical University, Guangzhou, People’s Republic of China, following the rules relating to the ethics on experimental animals. SuperScript^® II reverse transcriptase was purchased from Invitrogen (Grand Island, New York, USA). PrimeSTAR^® HS, restriction endonuclease, and a DNA Ligation Kit were purchased from Takara (Dalian, China). The pGEX4T-1 vector, TOP10, and BL21 (DE3) competent cells were from TIANGEN Biotech (Beijing, China). PageRuler Prest Protein Ladder was from Fermentas (Ontario, Canada). Trizol and isopropyl-β-d-thiogalactoside (IPTG) were from Sigma-Aldrich (St. Louis, Missouri, USA). Glutathione sepharose high-performance (GSH) beads were from BEAVER Nano (Suzhou, China). Goat anti-rabbit IgG-HRP antibody was from Santa Cruz Biotechnology (Dallas, Texas, USA). Alexa fluor 594 goat anti-rabbit IgG (H+L) secondary antibody conjugate was from Life Technologies (Grand Island). Bicinchoninic acid assay (BCA) protein assay kit was from Thermo Scientific (Waltham, Massachusetts, USA). Centrifugal filter units (30 kDa) were from Merck Millipore (Bedford, Massachusetts, USA).

Xiao B., Kuang Z., Zhan Y., Chen D., Gao Y., Li M., Luo S, & Hao W. (2016). A Novel Polyclonal Antiserum against Toxoplasma gondii Sodium Hydrogen Exchanger 1. The Korean Journal of Parasitology, 54(1), 21-29.

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