Tcep hcl

Manufactured by Thermo Fisher Scientific

Sourced in United States

TCEP-HCl is a reducing agent used in biochemical applications. It functions to reduce disulfide bonds in proteins and other biomolecules.

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Lab products found in correlation

Trypsin, by Promega (3 mentions) Madli tof ms microflex lt, by Bruker (2 mentions) Capture select hu lc kappa and lc lambda affinity resin, by Thermo Fisher Scientific (2 mentions) Sulfo smcc, by Thermo Fisher Scientific (2 mentions) Endoproteinase lys c, by Promega (2 mentions) Glu c, by Promega (2 mentions) Synergy 2 microplate reader, by Agilent Technologies (1 mentions) Gsh glo glutathione assay, by Promega (1 mentions) Trypan blue solution, by Merck Group (1 mentions) Cgrp8 37, by Bio-Techne (1 mentions) Carbenoxolone, by Merck Group (1 mentions) Dl tboa, by Bio-Techne (1 mentions) D apv, by Bio-Techne (1 mentions) Tris 2 carboxyethyl phosphine hydrochloride, by Thermo Fisher Scientific (1 mentions) Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions) Amylose resin, by New England Biolabs (1 mentions) Freestyle max reagent, by Thermo Fisher Scientific (1 mentions) Pmal c8, by Anatrace (1 mentions) Allyl isothiocyanate, by Merck Group (1 mentions) Bio beads sm 2, by Bio-Rad (1 mentions)

Spelling variants of this product

Tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl) (6 citations)

20 protocols using tcep hcl

Detecting Monoclonal Abnormalities by MALDI-TOF MS

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Example 10

Methods. Five hundred fifty six (556) serum samples that had been previously analyzed by routine clinical PEL/IFE testing were evaluated by MADLI-TOF MS (Microflex LT, Bruker Daltonics). Prior to analysis, intact immunoglobulins were isolated from serum with Capture Select™ (Hu)LC-kappa and LC-lambda affinity resin (Life Technologies) and reduced with tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl, Thermo Scientific). Purified samples were prepared for MALDI-TOF analysis using dried droplet method and α-cyano-4-hydroxycinnamic acid as matrix. Mass analysis was performed in positive ion mode with summation of 500 laser shots.

Results. For spectral analysis, the ion distribution of the MH+1 and MH+2 charge states of the light chain were compared to the spectrum of normal serum. Any monoclonal abnormalities were distinguished from the normal pattern. Of the 556 samples assayed, abnormal patterns were identified in 406 of 421 samples (96%) that were positive by IFE. Abnormalities were also noted in 23 of 126 samples (18%) that were negative by IFE. Of the 9 samples that were indeterminate by IFE, abnormalities were noted in 2.

US11604196B2. Isotyping immunoglobulins using accurate molecular mass (2023-03-14). Mayo Foundation for Medical Education and Research [US]. Inventors: David L. Murray [US], David R. Barnidge [US], Surendra Dasari [US], John R. Mills [US].

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Quantification of Cellular Glutathione Levels

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Total extracellular and intracellular glutathione (GSH + GSSG) concentrations were determined using the GSH-Glo glutathione assay (Promega, Madison, WI) per manufacturer’s instruction. Media was collected for analysis of the extracellular GSH ([GSH]_e) and GSH-Glo reaction buffer was added to the cells for analysis of the intracellular GSH ([GSH]_i). Additional reaction buffer was made per manufacturer’s instructions and 200 µL was added per well as a dilution factor to keep relative light units within the dynamic range of the standards. To determine total GSH levels, glutathione disulfide (GSSG) within the samples was converted to GSH with the reducing agent TCEP-HCl (final concentration = 1 mM; 10 min; 25°C; Thermo Scientific; Waltham, MA). Luciferase activity was measured using an Optocomp II luminometer (MGM Instruments) or a Synergy² microplate reader (BioTek, Winooski, VT). Total intracellular or extracellular GSH were normalized to standards prepared in GSH-Glo reaction buffer and MS containing 0.1% fatty acid free BSA, respectively. Extracellular GSSG was calculated based on the equation GSSG = (total GSH – reduced GSH) / 2. [GSH]_i were normalized to cellular protein. Standards were linear over the range of 0–5 µM.

He Y., Jackman N.A., L.Thorn T., Vought V.E, & Hewett S.J. (2015). Interleukin-1β protects astrocytes against oxidant-induced injury via an NFκB-dependent upregulation of glutathione synthesis. Glia, 63(9), 1568-1580.

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Trigeminal Ganglion Injections: Detailed Procedure

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The technique for trigeminal ganglion injections followed those detailed by Neubert et al.²⁶ Briefly, the head of an anesthetized animal was stabilized in one hand and the rostral portion of the zygomatic process of the maxillary bone palpated. A sterile 25 gauge ×20 mm needle was then inserted medial to the zygomatic process through the infraorbital foramen. Drugs dissolved in saline and 0.2% Trypan Blue solution (Sigma # T8154) were then injected (7 µl) over 1 min using a Hamilton syringe. The needle remained in the foramen for 5 min and was then slowly removed. Trypan Blue served as verification of successful drug injection into the ganglia and animals in which the dye was not visible or in which dye was observed outside the ganglion following blood flow measurements were excluded from analysis. Injected drugs included carbenoxolone (Sigma #C4790), CGRP₈_-₃₇ (Tocris #1169), DL-TBOA (Tocris #1223), D-APV (Tocris #0106), and A-317491 (Santa Cruz #sc-300144). Saline and CGRP₈_-₃₇ inactivated with the reducing agent TCEP-HCL (Thermoscientific #20490) were used as controls. Agents were injected under anesthesia 2 h before blood flow measurements.

Zhang L., Kunkler P.E., Knopp K.L., Oxford G.S, & Hurley J.H. (2019). Role of intraganglionic transmission in the trigeminovascular pathway. Molecular Pain, 15, 1744806919836570.

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Bacterial Aptamer Biosensor Development

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Aptamers are custom-designed to selectively bind with the DNA of Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus, and an aptamer for total bacteria detections as mentioned below (Figure 1) was purchased from Genotech, Daejeon, South Korea.
To prepare the DNA probe for immobilization on the Au electrode surface, under optimized conditions, a concentration of 1 mM was prepared. Next, the diluted DNA probe was mixed with 10 mM Tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl, Thermo Scientific™, Waltham, MA, USA) in the dark and incubated for 1 h. This step is crucial for reducing the disulfide bonds present in the thiol-modified aptamer end.

Wu T., Yagati A.K, & Min J. (2023). Electrochemical Detection of Different Foodborne Bacteria for Point-of-Care Applications. Biosensors, 13(6), 641.

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Methamphetamine Immunization Protocol

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(+)-Methamphetamine hydrochloride (METH) and the tritiated form of METH ([³H]-METH) were obtained from the NIDA Drug Supply Program. GLA-SE adjuvant was from Immune Design Corp. (Seattle, WA). Sulfo-SMCC (Sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate) crosslinker and TCEP-HCl (Tris(2-carboxyethyl)phosphine hydrochloride) were purchased from Thermo Fisher Scientific Inc. (Pierce, Rockford, IL). Vacmune®, two purified monomers of keyhole limpet hemocyanin (KLH), was obtained from Biosyn Corp. (Carlsbad, CA).

Stevens M.W., Rüedi-Bettschen D., Gunnell M.G., Tawney R., West C.M, & Owens S.M. (2019). Antibody production and pharmacokinetics of METH in rats following vaccination with the METH vaccine, IXT-v100, adjuvanted with GLA-SE. Drug and alcohol dependence, 204, 107484.

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Immobilization of Thiol-modified ssDNA on Mal-T80A20

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A 3′-thiol-C3-modified (disulphide) single-stranded DNA oligomer (ssDNA, Integrated DNA Technologies, 73 nucleotides, see Supplementary Information) was added in 10-fold excess to mal-T80A20 by mixing (10 µL) of 20 µM mal-T80A20 (200 pmoles) with 2 nmoles of ssDNA in 1 mM PBS, (5 mM) TCEP·HCl (ThermoFisher Scientific cat. #:20490) in a (100 µL) reaction volume. The reaction was mixed for at least 30 mins and washed with 50 kDa MWCO centrifugation filtering (x5 washes, 2 k × g, 10 mins, 3.5 mL wash 1 mM PBS). Wash effluent was collected and inspected for unbound ssDNA by measuring absorbance at 260 nm using NanoDrop UV-visible spectroscopy. Washes were continued until there was no unbound DNA detected in the wash effluent for three consecutive washes.

Edgington R., Spillane K.M., Papageorgiou G., Wray W., Ishiwata H., Labarca M., Leal-Ortiz S., Reid G., Webb M., Foord J., Melosh N, & Schaefer A.T. (2018). Functionalisation of Detonation Nanodiamond for Monodispersed, Soluble DNA-Nanodiamond Conjugates Using Mixed Silane Bead-Assisted Sonication Disintegration. Scientific Reports, 8, 728.

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Synthesis of Genentech GNE Analogs

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GNE551:5-amino-1-(4-bromo-2-fluorobenzyl)-N-(2,5-dimethoxyphenyl)-1H-1,2,3-triazole-4-carboxamide, GNE551 analogs, WaTx, A-96079, AMG0902, and GNE235 were synthesized at Genentech. The following reagents were obtained commercially: allyl isothiocyanate (Sigma-Aldrich); Hank’s Balanced Salt Solution, FreeStyle Max Reagent, Calcium assay kit, and TCEP-HCl (Thermo Fisher); Facade®-EM (Avanti); Bio-Beads SM-2 (Bio-Rad); PMAL-C8 (Anatrace); and Amylose resin (New England Biolabs).

Liu C., Reese R., Vu S., Rougé L., Shields S.D., Kakiuchi-Kiyota S., Chen H., Johnson K., Shi Y.P., Chernov-Rogan T., Greiner D.M., Kohli P.B., Hackos D., Brillantes B., Tam C., Li T., Wang J., Safina B., Magnuson S., Volgraf M., Payandeh J., Zheng J., Rohou A, & Chen J. (2020). A non-covalent ligand reveals biased agonism of the TRPA1 ion channel. Neuron, 109(2), 273-284.e4.

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Proteomic Analysis of Dendritic Cells

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Equal protein amounts (50 μg) from total DCs lysates (B6 and Ob/Ob), were reduced in 20 mM TCEP.HCl (Thermo Scientific), 50 mM ammonium bicarbonate buffer, at pH 8.5 containing 8 M urea for 35 min at room temperature. The reduced proteins were alkylated with 100 mM iodoacetamide solution, for 50 min at room temperature in the dark. Three different enzymes were used for “in solution” digestion in 50 mM ammonium bicarbonate buffer, pH 8.5, for 18 h, at 37 °C: endoproteinase Lys-C (1:50 enzyme: protein ratio); trypsin (1:20 enzyme: protein ratio) and Glu-C (1:10 enzyme: protein ratio) (sequencing grade Promega, Madison, WI, USA. Peptides mixture, extracted from all enzymatic digestions, were desalted on C18 Prep clean columns before high resolution liquid chromatography tandem mass spectrometry (LC-MS/MS).

Clement C.C., Nanaware P.P., Yamazaki T., Negroni M.P., Ramesh K., Morozova K., Thangaswamy S., Graves A., Kim H.J., Li T.W., Vigano’ M., Soni R.K., Gadina M., Tse H.Y., Galluzzi L., Roche P.A., Denzin L.K., Stern L.J, & Santambrogio L. (2021). Pleiotropic consequences of metabolic stress on the major histocompatibility complex class II molecule antigen processing and presentation machinery. Immunity.

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Quantitative Proteomics of Lymph Samples

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Total protein concentration for pre- and post-nodal lymph samples was determined using the Bradford micro assay. Equal protein amounts (20 μg), prepared in technical triplicates, were reduced in 15 mM TCEP.HCl (Thermo Scientific), 50 mM ammonium bicarbonate buffer, at pH 8.5, for 35 min at room temperature. The reduced proteins were further alkylated with 55 mM iodoacetamide solution, for 50 min at room temperature. Three different enzymes were used for “in solution” digestion in 50 mM ammonium bicarbonate buffer, pH 8.5, for 18 h, at 37 °C: endoproteinase Lys-C (1:50 enzyme: protein ratio); trypsin (1:20 enzyme: protein ratio) and Glu-C (1:10 enzyme: protein ratio) (sequencing grade Promega, Madison, WI, USA). The peptides mixture, extracted from all enzymatic digestions, were desalted on C18 Prep clean columns before high resolution liquid chromatography tandem mass spectrometry (LC-MS/MS).

Clement C.C., Wang W., Dzieciatkowska M., Cortese M., Hansen K.C., Becerra A., Thangaswamy S., Nizamutdinova I., Moon J.Y., Stern L.J., Gashev A.A., Zawieja D, & Santambrogio L. (2018). Quantitative Profiling of the Lymph Node Clearance Capacity. Scientific Reports, 8, 11253.

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Site-Specific Protein Dimerization via TCEP-Mediated Ligation

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KR13 (1; 2.084 mg, 2.5 equiv) was dissolved into 1000 μL of degassed phosphate buffer (50 mM, pH 6.6) prior to the addition of 0.574 mg of tris(2-carboxyethyl)phosphine hydrochloride (TCEP·HCl, 5 equiv; Thermo Fisher Scientific) to the reaction vessel. Compound 1 was reduced with TCEP for 30 min under vacuum and nitrogen purging. Bis-Mal-dPeg₃ (400 μL, 0.522 g; Quanta Biodesign Limited) dissolved into 1000 μL of ACN (1 equiv) was added dropwise to the reaction vessel. The ligation process was monitored by analytical HPLC (Phenomenex Luna 5 μ C18, 100 Å, 250 mm × 4.6 mm, 1 mL/min) over a 30–95% ACN gradient, as the reaction mixture was stirred under N₂ at 37 °C. After 2 h, another 200 μL of Bis-Mal-dPeg was added to the reaction, and the reaction continued for 3 h. Peptide purity and mass of the final ligation product, KR13 dimer (1a), were confirmed using an analytical HPLC (Phenomenex C18) and MALDI-TOF mass spectrophotometry, respectively.

Bailey L.D., Sundaram R.V., Li H., Duffy C., Aneja R., Bastian A.R., Holmes A.P., Kamanna K., Rashad A.A, & Chaiken I. (2015). Disulfide Sensitivity in the Env Protein Underlies Lytic Inactivation of HIV-1 by Peptide Triazole Thiols. ACS chemical biology, 10(12), 2861-2873.

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