Ultra-pure HPLC water was from VWR (Fontenay-sous-Bois, France), HPLC gradient grade acetonitrile, nonafluoropentanoic acid (NFPA) 97%, hydrochloric acid (HCl) 37%, sodium borohydride (NaBH4), boric acid (H3BO3) and sodium hydroxide (NaOH), trichloroacetic acid (TCA), lysine (Lys), bovine serum albumin (BSA fraction V), glyoxylic acid and sodium cyanoborohydride (NaBH3CN) were all obtained from Sigma–Aldrich (Saint-Quentin-Fallavier, France). The labeled internal standard (15N2)-Lys was purchased from CortecNet (Voisins-le-Bretonneux, France), while CML, (D2)-CML, (D4)-CML, CEL, and (D4)-CEL were all from Polypeptide Group Laboratories (Strasbourg, France). A 0.1 M NaBH4 solution was made up of a 0.2 M borate buffer comprised of (H3BO3) and NaOH (pH = 9.5).
Sodium cyanoborohydride nabh3cn
Manufactured by Merck Group
Sourced in France, United States
Sodium cyanoborohydride (NaBH3CN) is a reducing agent used in various laboratory applications. It is a white, crystalline solid that is soluble in water and certain organic solvents. The core function of sodium cyanoborohydride is to selectively reduce imines and aldehydes to secondary amines and alcohols, respectively, without affecting other functional groups in the molecule.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Hplc gradient grade acetonitrile, by Avantor (1 mentions)
Nonafluoropentanoic acid nfpa, by Merck Group (1 mentions)
Sodium borohydride nabh4, by Merck Group (1 mentions)
Boric acid h3bo3, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Restriction endonuclease, by Takara Bio (1 mentions)
Prime startm hs dna polymerase, by Takara Bio (1 mentions)
Standard gag unsaturated disaccharides, by Iduron (1 mentions)
Acetonitrile of hplc grade, by Merck Group (1 mentions)
Formaldehyde ch2o, by Merck Group (1 mentions)
Milli q gradient water purification system, by Merck Group (1 mentions)
Sequencing grade modified trypsin, by Worthington (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Ammonia solution, by Fujifilm (1 mentions)
Formic acid, by Fujifilm (1 mentions)
Trifluoroacetic acid, by Thermo Fisher Scientific (1 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
12 protocols using sodium cyanoborohydride nabh3cn
1
Quantification of Glycation Adducts in Proteins
2
Enzymatic Chondroitin Sulfate Characterization
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The strains and plasmids used in this study are listed in Table 1 . PrimeSTARTM HS DNA polymerase, restriction endonucleases and other genetic engineering enzymes were purchased from Takara Inc. (Dalian, China). A genomic DNA extraction kit was purchased from Tiangen Biotech Co., Ltd. (Beijing, China). Competent cells and reagents for site-directed mutagenesis were obtained from TransGen Biotech Co., Ltd. (Beijing, China). Reagents for protein crystallization were purchased from Hampton Research (Aliso Viejo, United States). Standard GAG unsaturated disaccharides were purchased from Iduron (Manchester, United Kingdom). DS from porcine skin were purchased from Seikagaku Corp. (Tokyo, Japan). CS-A from bovine cartilage, chondroitinase ABC (CSase ABC) (EC 4.2.2.4), 2-aminobenzamide (2-AB), sodium borohydride and sodium cyanoborohydride (NaBH3CN) were purchased from Sigma-Aldrich Inc. The hexasaccharide ΔA-A-A [Δ4,5HexUAβ1-3GalNAc(4S)β1-4Gl cUAβ1-3GalNAc(4S)β1-4GlcUAβ1-3GalNAc(4S)] was prepared from CS-A as described below.
3
Bovine Milk Protein Quantitation
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BSA was used
as the standard for protein quantitation and purchased from Sigma-Aldrich
(St. Louis, MO). Sodium bicarbonate (NaHCO3), dithiotheritol
(DTT), iodoacetamide (IAA), ammonium hydroxide, sodium cyanoborohydride
(NaBH3CN), formaldehyde (CH2O), and formaldehyde-isotope
(13CD2O) were from Sigma-Aldrich (St. Louis,
MO) as well. Sequencing grade modified trypsin was from Worthington
Biochemical Corporation (Freehold, NJ). Formic acid and acetonitrile
of HPLC grade were obtained from Merck (Darmstadt, Germany). Ultrapure
water was obtained by a Milli-Q Gradient water purification system
(Millipore, Bedford, MA). Raw bovine milk samples were provided by
Shiyun Lai in Beingmate Research Institute.
as the standard for protein quantitation and purchased from Sigma-Aldrich
(St. Louis, MO). Sodium bicarbonate (NaHCO3), dithiotheritol
(DTT), iodoacetamide (IAA), ammonium hydroxide, sodium cyanoborohydride
(NaBH3CN), formaldehyde (CH2O), and formaldehyde-isotope
(13CD2O) were from Sigma-Aldrich (St. Louis,
MO) as well. Sequencing grade modified trypsin was from Worthington
Biochemical Corporation (Freehold, NJ). Formic acid and acetonitrile
of HPLC grade were obtained from Merck (Darmstadt, Germany). Ultrapure
water was obtained by a Milli-Q Gradient water purification system
(Millipore, Bedford, MA). Raw bovine milk samples were provided by
Shiyun Lai in Beingmate Research Institute.
4
Dimethyl Labeling of Peptides
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The stable isotope dimethyl labeling involves the formation of a Schiff base via the reaction of formaldehyde with the primary amines, which are then reduced by cyanoborohydride67 (link). The digested peptides of two controls, MCM2-overexpressed and MCM2-silenced samples were dried using a centrifugal evaporator and reconstituted separately with 100 mM TEABC. Each control sample was labeled with 4% formaldehyde-H2 (37% Formaldehyde solution; Sigma-Aldrich). MCM2-overexpressed and MCM2-silenced samples were labeled with 4% formaldehyde-D2 (20% Formaldehyde-13C, d2 solution; Sigma-Aldrich) separately. 4 μL of 600 mM sodium cyanoborohydride (NaBH3CN, Sigma-Aldrich) were then added to each sample solution and incubated for 1 h at room temperature. The reaction was inactivated by adding 16 μl of 1% (v/v) ammonia solution (WAKO) and acidified the peptides using 10% (v/v) formic acid (WAKO) to a pH < 3. The H2-labeled A549 control and H1299 control were combined with the D2 labeled MCM2-overexpressed and MCM2-silenced samples at 1:1 ratio, respectively. These two combined mixtures were subjected to desalting as described previously.
5
Peptide Synthesis Reagents Procurement
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Amino acids, N,N′-diisopropylethylamine (DIEA), N,N′-Disuccinimidyl carbonate (DSC), Fmoc chloride, (1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU), chloro-N,N,N′,N′-tetramethylformamidinium hexafluorophospate (TCFH) and triphosgene (BTC) were purchased from Chem Impex Int’l, Inc. Reagents such as piperidine, lithium chloride, sodium cyanoborohydride (NaBH3CN), triisopropylsilane (TIPS), sodium acetate-d3, acetic acid-d4, and D2O were purchased from Sigma Aldrich. Reagents such as hydrazine monohydrate and 2,4,6-collidine were purchased from Alfa Aesar. Trifluoroacetic acid, glacial acetic acid, sodium bicarbonate, and solvents were purchased from Fisher. Reagents including HBTU and PyBOP were purchased from Oakwood Chemical. 1-Methylimidazole (NMI) and 1-chloro-N,N,2-trimethylpropenylamine (Ghosez’s Reagent) were purchased from Acros Organics.
6
Fluorescent Nanodiamonds for Biomedical Applications
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Fluorescent nanodiamonds (FNDs, carboxylated, 100 nm, 1 mg mL−1) in DI water) were obtained from Adamas Nano (Raleigh, NC), sodium hyaluronate (HA, 200 kDa) from Lifecore Co. (Chaska, MN) and N-hydroxysulfosuccinimide (sulfo-NHS) from Georgiachem. 1,4-Diaminobutane (DAB) and sodium cyanoborohydride (NaBH3CN) were purchased from Sigma Aldrich (Seoul, Korea). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and 5-aminofluorescein (FTIC) were purchased from Tokyo Chemical Industry (Tokyo, Japan). Dulbecco's modified Eagle's medium-high glucose (DMEM), fetal bovine serum (FBS), antibiotics, and phosphate buffered saline (PBS, pH 7.4) were obtained from Invitrogen (Carlsbad, CA). Cell counting kit-8 (CCK-8) was obtained from DoGenBio Co. Human liver epithelial cell line (FL83B), human liver cancer cell line (HepG2), adenocarcinomic human alveolar basal epithelial cells (A549) and human embryonic kidney 293 cells (HEK293) were purchased from Korean Cell Line Bank (Seoul, Korea). 6 week-old female balb/c nude mice were purchased at the Orient Bio (Seongnam, Korea). Alfalfa free feed (AIN-93G) for mouse was purchased from Saeronbio Co. (Uiwang, Korea). All animal experiments followed the guidelines for Care and Use of Laboratory Animals of the Pohang University of Science and Technology (POSTECH) and approved by the Institutional Animal Care and Use Committee (IACUC).
7
Targeted Drug Delivery Nanoparticle Synthesis
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Polyethylene glycol monomethyl ether (mPEG), L-phenylalanine (L-Phe), L-cysteine (L-Cys), and deuterated trifluoroacetic acid (TFA-d) were purchased from Sigma-Aldrich (Shanghai, PR China). The STP, DOX hydrochloride (DOX HCl), and GSH were obtained from Gill Biochemical Co., Ltd. (Shanghai, PR China). Cell culture products, including Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS), were provided by Gibco (USA). Penicillin and streptomycin were obtained from Huabei Pharmaceutical Co., Ltd. (Hebei, PR China). Sodium cyanoborohydride (NaBH3CN), 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (Shanghai, P. China). Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) kit was purchased from Roche Company (Mannheim, Germany). The purified deionized water was prepared by the Milli-Q plus system (Millipore Co., Billerica, MA, USA).
8
N-glycan Labeling and Purification
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The A2G0 N-glycan was purchased from Prozyme (Hayward, CA). Endoglycosidase Endo S was purchased from New England Biolabs (Ipswich, MA, USA). Natural abundance aniline and [13C6] aniline, sodium cyanoborohydride (NaBH3CN), were purchased from Sigma Aldrich (Milwaukee, WI, USA). Hypersep Hypercarb PGC SPE cartridges (25 mg, 1 mL) and Pierce BCA Protein Assay Kit were purchased from ThermoFisher Scientific (Waltham, MA, USA). Creatinine Assay Kit was purchased from Sigma (St. Louis, MO, USA). The ganglioside standards (GM1, GA1, GM2, GA2 and GM3) were all purchased from Enzo Life Sciences (Farmingdale, NY, USA) while the BMP(14:0) phospholipid was purchased from Avanti Polar Lipids (Alabaster, AL, USA). The 10kDa centrifugal filters were from VWR (Radnor, PA, USA). All other chemicals were of reagent or LC-MS grade for chromatography systems.
9
Surface Antigen and Antibody Immobilization
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HBsAg (Human), HBV surface antibody (HBsAb), and P16INK4A antigen (Human) were purchased from Proteintech Inc. (Rosemont, IL, USA). The antigens and antibodies were stored in −20 °C environment after dilution. (3-aminopropyl)triethoxysilane [APTES; H2N(CH2)3Si(OC2H5)3, MW: 221.37 g/mol], analytical-grade ethanol (C2H5OH, absolute ≥99.8%), glutaraldehyde [GA; OHC(CH2)3CHO; MW: 100.12 g/mol], Bis-tris propane {BTP; CH2[CH2NHC(CH2OH)3]2, molecular weight (MW): 282.34 g/mol}, sodium cyanoborohydride (NaBH3CN; 95%, MW: 62.84 g/mol), and Tris hydrochloride [Tris HCl, (HOCH2)3CNH2•HCl, MW:157.6] were obtained from Sigma-Aldrich (St. Louis, MO, USA).
10
Analytical Glycan Quantification Protocol
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Glc4 (d -glucopyranosyl maltotriose, Glcα1-6Glcα1-4Glcα1-4Glc) was from TRC, Toronto, Canada). 13C6 hexose tetrasaccharide internal standard (6C13-labeled mixture of Glc4 and isomaltose-isomaltose, Glcα1-6Glcα1-4Glcα1-6Glc, IS) was a generous gift of Dr. S. Young (Duke Biochemical Genetics Laboratory, Durham, NC, USA). Sodium cyanoborohydride (NaBH3CN) and butyl-4-aminobenzoate (BAB) were from Sigma-Aldrich® (Saint Quentin-Fallavier, France). Acetonitrile (ACN) for HPLC, methanol and acetic acid were from Carlo-Erba (Val de Reuil, France). HPLC grade water was from Ecotainer Aqua B.Braun© (Melsungen, Germany). Bond Elut C18 50 mg, 1 mL solid phase extraction (SPE) columns were used with a Vac Elut 20 system, all from Varian (Agilent, Santa Clara, CA). Reverse phase LC was performed on a Uptisphere 3BioP2 C18 column (100 × 2.1 mm, 3 μm, ref. UP3BP2#10QS, Interchrom©, Interchim, Montluçon, France).
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