Human cDNAs coding for FMNL2 (Q96PY5-3) and FMNL3 (Q8IVF7-3) were purchased from Promega (Madison, WI, USA). ECL Prime Western Blotting Detection Reagent was sourced from GE Healthcare (Amersham, UK). The dye terminator cycle sequencing kit, Lipofectamine LTX with PLUS reagent and Hoechst 33342 were obtained from Life Technologies Corp. (Carlsbad, CA, USA). Anti-FLAG monoclonal antibody and anti-mouse IgG-FITC antibody were obtained from Sigma (St. Louis, MO, USA). Tetradec-13-ynoic acid (Alk-Myr) was sourced from Cayman Chemical Co. (Ann Arbor, MI, USA). 5-TAMRA Azide (Az-TAMRA) was purchased from Click Chemistry Tools (Scottsdale, AZ, USA). Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA) were obtained from Sigma (St. Louis, MO, USA). Protein G-HRP conjugate was sourced from Bio-Rad (Hercules, CA, USA). HRP-conjugated anti-mouse IgG was purchased from Cell Signaling Technology (Danvers, MA, USA). The other reagents used were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) or Daiichi Pure Chemicals Co., Ltd. (Tokyo, Japan), and were of analytical or DNA grade.
Tris 2 carboxyethyl phosphine hydrochloride tcep
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) is a reducing agent commonly used in biochemistry and molecular biology applications. It is a water-soluble, odorless, and stable compound that efficiently reduces disulfide bonds in proteins and peptides.
Automatically generated - may contain errors
Lab products found in correlation
Iodoacetamide, by Merck Group (5 mentions)
Sodium chloride (nacl), by Merck Group (4 mentions)
Hepes, by Merck Group (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Formic acid, by Thermo Fisher Scientific (4 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (4 mentions)
6 mercaptohexanol, by Merck Group (3 mentions)
6 mercapto 1 hexanol, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Potassium chloride (kcl), by Merck Group (3 mentions)
Plasmin, by Enzyme Research (3 mentions)
Potassium chloride (kcl), by Thermo Fisher Scientific (3 mentions)
Ultramicro spin column, by Nest Group (3 mentions)
Anti flag monoclonal antibody, by Merck Group (2 mentions)
Anti mouse igg fitc antibody, by Merck Group (2 mentions)
Tris 1 benzyl 1h 1 2 3 triazol 4 yl methyl amine tbta, by Merck Group (2 mentions)
Protein g hrp conjugate, by Bio-Rad (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Fused silica capillary tubing, by Molex (2 mentions)
Bovine thyroglobulin btg, by Thermo Fisher Scientific (2 mentions)
77 protocols using tris 2 carboxyethyl phosphine hydrochloride tcep
1
Cloning and Detection of FMNL2 and FMNL3
2
Nanoparticle Formulation for siOPN Delivery
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PLGA (50:50, ester terminated, MW of 30–60 kDa), tris-EDTA buffer (TE buffer, RNase-free, 10 mM Tris-HCl, 1 mM EDTA), PEI (branched, 800 Da), poly(vinyl alcohol) (PVA, 30–70 kDa), N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), 1-Hydroxybenzotriazole (HOBt), and Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) were purchased from Sigma-Aldrich (Rehovot, Israel). PLGA 50:50, acid terminated, MW of 50–60 kDa was purchased from Lakeshore Biomaterials (Birmingham, AL, USA). PLGA-PEG copolymer (RGP d 50105, PLGA, 45 kDa, and PEG, 5 kDa) was purchased from Boehringer Ingelheim (Ingelheim, Germany). The heterobifunctional PEG, amine-PEG-maleimide (NH2-PEG-MAL; PEG, 2 kDa) was purchased from Creative PEGWorks (Chapel Hill, NC, USA). siOPN (described in [22 (link),23 (link)]) was custom synthesized by Thermo Fisher Scientific (Waltham, MA, USA). The navigator peptide, ApoB-P (a 25 AA sequence: SVKAQWKKNKHRHGCGRLTRKRGLK [30 ]; MW of ~3 kDa) was purchased from CASLO ApS (Lyngby, Denmark). Mannitol and organic solvents were obtained from J.T. Baker Chemicals (Radnor, PA, USA). Tissue culture reagents and phosphate buffered saline (PBS) were purchased from Biological Industries (Beit-Haemek, Israel).
3
Synthesis and Functionalization of Gold Nanoparticles
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Gold chloride trihydrate (HAuCl4·3H2O), sodium citrate, sodium borodeuteride (NaBH4) were obtained from Aladdin Biochemical Technology Co. Ltd. (Shanghai, China). Ti3C2 dispersion liquid was obtained from Jiangsu XFNANO Materials Tech, Co. Ltd. (Nanjing, China). Tris (4,4′-dicarboxylic acid-2,2′-bipyridyl) ruthenium (II) dicbloride (Ru(dcbpy)3Cl2) was obtained from Suna Tech Inc. (Suzhou, China). Tris(2-carboxyethyl) phosphine hydrochloride (TCEP) and 6-mercaptohexanol (MCH) were gained from Sigma-Aldrich (St Louis, MO, USA). The purified DNA sequences (Table S1 ) were synthesized by Genscript Bio-technology Co. Ltd. (Nanjing, China).
4
Targeted Lipid Nanoparticle Delivery
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1,2-Distearoyl-sn-glycero-2-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide (polyethylene glycol)-2000] (DSPE-PEG(2000) maleimide), and 1,2-distearoyl-sn-glycero-3-phosphoetha-nolamine-N-[carboxy (polyethylene glycol)-2000] (DSPE-PEG(2000) carboxylic acid) were purchased from Avanti Polar Lipids (Alabaster, AL, USA). N-Succinimidyl-3-(2-pyridyldithio) propionate (SPDP), cholesterol, atosiban, and tris(2-carboxyethyl) phosphine hydrochloride (TCEP) were from Sigma-Aldrich Co. (St Louis, MO, USA). 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), CBQCA protein assay kit and N-hydroxysuccinimide (NHS) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Rabbit IgG antibody and anti-OTR monoclonal antibody (ab181077) were obtained from Abcam (Cambridge, UK). PD-10 column was from VWR International (Radnor, PA, USA). All other chemicals and solvents were of at least analytical grade.
5
Glycoproteome Analysis Using AminoxyTMT
6
Aptamer-Functionalized AuNP Sensor Fabrication
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The aptamer (H8DAE) was synthesized, characterized using MST, and purchased from Base Pair Biotechnologies (Pearland, Texas). The NC membrane (pore size of ) and blotting absorbent filter paper (# 1703960) were purchased from BioRad (Hercules, California), and the pure cellulose chromatography paper (Cat. No. 05714-1) was acquired from Fisher Scientific (Waltham, Massachusetts). For the synthesis of the citrate-capped AuNPs, gold (III) chloride ( ) and sodium citrate were obtained from Sigma-Aldrich (St. Louis, Missouri). Before conjugation of the thiolated aptamer, the disulfide bonds were reduced using tris(2-carboxyethyl)phosphine hydrochloride (TCEP) purchased from Sigma-Aldrich (St. Louis, Missouri). The immobilization of the AuNPs on the nitrocellulose membrane was performed using streptavidin, which was purchased from VWR (Radnor, Pennsylvania) and a PEGylated-biotin linker was obtained from ThermoFisher Scientific (Waltham, Massachusetts) and modified accordingly. All experiments were performed in phosphate-buffered saline ( PBS) at pH 7.4 and Raman measurements were taken using an ID Raman mini 2.0 (Metrohm—formally purchased from Ocean Optics, Switzerland).
7
Proteomic Analysis of Cancer Cell Lines
8
Protein Secondary Structure Analysis by CD Spectroscopy
9
Duplex DNA Formation and Characterization
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The oligomers for the duplex DNA formed by ssDNA (5′-S-S-(C6H12)-TTT ACC TTT ATT-3′) and cDNA (3′-(MB)-AAA TGG AAA TAA CC-5′) were synthesized by and purchased from Integrated DNA Technologies* (Coralville, IA, USA) and Biosearch Technologies* (Novato, CA, USA), respectively. Dimethyl sulfoxide (DMSO), diminazene aceturate (DMZ), 6-mercaptohexanol, proflavine hemisulfate salt hydrate, sodium phosphate dibasic, sodium phosphate monobasic, magnesium chloride heptahydrate, sodium chloride, and tris-(2-carboxyethyl) phosphine hydrochloride (TCEP) were purchased from Sigma-Aldrich* (St. Louis, MO, USA). All chemical reagents were of analytical grade or higher. Stock solutions of proflavine and DMZ were prepared at 14 mmol/L in DMSO and then diluted to 140 µmol/L in PBS buffer (10 mmol/L phosphate-buffered saline, pH 7.4) with 100 mmol/L NaCl. Deionized ultra-filtered (DIUF) water (18.2 MΩ·cm) was used in all preparations. Fused silica wafers were purchased from University Wafers* (Boston, MA, USA).
10
Rituximab and Adalimumab Formulation Preparation
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A volume of 6 ml of a rituximab formulation with a concentration of 10 mg/mL (MabThera®, Hoffmann-La Roche, N7025B04 exp. 2/2017; H0102B06 exp. 05/2014) was mixed with 0.72 g glucose dissolved in 2 mL H2O. For adalimumab (Humira®, AbbVie; exp. 2016), 2 mL of formulation buffer containing 10 mg/mL was mixed with 8 mL of a 0.625 M solution of glucose. The samples were incubated at 40°C for 7 days. Dialysis, lyophilization and dissolving under denaturing conditions followed the general procedure resulting in a final concentration of 30 mg/mL corresponding to a concentration of 0.2 mmol/L. The disulfide bridges were reduced by adding tris(2-carboxyethyl)phosphine hydrochloride (TCEP; Sigma Aldrich) and heating to 60°C for 15 min.
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