Tetramethylethylenediamine temed

Manufactured by Thermo Fisher Scientific

Tetramethylethylenediamine (TEMED) is a chemical reagent used in various laboratory procedures, particularly in gel electrophoresis. It functions as a catalyst, accelerating the polymerization of acrylamide and other monomers to form a gel matrix. TEMED is commonly used in combination with ammonium persulfate to initiate and maintain the polymerization process.

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TEMED (34 citations) N,N,N',N'-tetramethylethylenediamine (TEMED) (4 citations) Tetramethylethylenediamine (3 citations) N,N,N′,N′-Tetramethylethylenediamine (TEMED; C6H16N2), (2 citations)

5 protocols using tetramethylethylenediamine temed

Capillary and Slab Gel Electrophoresis

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General procedure for preparing gel columns was followed. Briefly, 12% resolving gel was prepared by mixing 3.33 ml water, 2.5 mL Tris-HCl buffer (pH-8.8), 4 mL mixture of acrylamide and Bis-acrylamide (Bio-Rad) in ratio 29:1, 100 µL of 10% SDS, 60 µL Ammonium persulphate (APS) (Sigma Aldrich) and 6.5 µL of Tetramethylethylenediamine (TEMED) (Fisher Scientific). 6% stacking gel was prepared by adding 700 µL mixture of acrylamide: Bis-acrylamide in ratio 29:1, 675 µL of Tris-HCl buffer (pH-6.8) and 50 µL of 10% SDS. To this 25 µL of APS and 20 µL of TEMED were added. For capillary (Fisherbrand) preparation, resolving gel columns of 1.5 or 2.5 cm length were made by injecting 30 or 50 µL of resolving gel solution into the capillary tube, respectively. This was followed by injecting 10 µL of stacking gel solution to form stacking gel of 0.5 cm length. The standard PAGE slab was prepared using the above mentioned solutions to form resolving and stacking gels of 4 cm and 1 cm length.

Chaudhari S., Chaudhari K., Kim S., Khan F., Lee J, & Thundat T. (2017). Electrophoresis assisted time-of-flow mass spectrometry using hollow nanomechanical resonators. Scientific Reports, 7, 3535.

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Optimized CMC-MC Hydrogel Formulation

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The formulation of CMC-MC used in this study was optimized in a previous
investigation (Varma et al., 2018a (link)).
Methacrylated CMC and MC, dissolved in phosphate-buffered saline (PBS)
(Sigma-Aldrich) at 3% (w/v) macromer concentration each, were homogeneously
combined in both barrels of dual-barrel syringes (Pac-Dent, Brea, CA). Redox
initiators, ammonium persulfate (APS) (Sigma-Aldrich) and
N,N,N’,N’-tetramethylethylenediamine (TEMED) (Fisher Scientific,
Hampton, NH) were added to the separate barrels of the syringes at 20 mM each
and mixed with the polymer solution. Trypan Blue (Fisher Scientific) was added
to the polymer solution as well to facilitate enhanced visual detection of the
hydrogel post gelation. Following a 30-min incubation period in a 37°C
water bath to initiate MC gelation, the polymer mixtures were injected via a 20G
needle (Fisher Scientific) attached to a mixing tip (Pac-Dent) connected to the
syringe (Figure 1A)

Lin H.A., Varma D.M., Hom W.W., Cruz M.A., Nasser P.R., Phelps R.G., Iatridis J.C, & Nicoll S.B. (2019). Injectable Cellulose-Based Hydrogels as Nucleus Pulposus Replacements: Assessment of In Vitro Structural Stability, Ex Vivo Herniation Risk, and In Vivo Biocompatibility. Journal of the mechanical behavior of biomedical materials, 96, 204-213.

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Fluorescent Hydrogel Synthesis Protocol

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Acrylamide (AAm), N-hydroxysuccinamide, 4’Aminofluorescein isomer I, 5(6)-Carboxyfluorescein, Fluorescein disodium salt, FITC, 3-(trimethoxysilyl) propyl methacrylate and Ethylene glycol were purchased from Acros Organics. Pyruvic acid and 3-aminopropyl methacrylamide hydrochloride (3AP) were purchased from Sigma-Aldrich. Tetramethylethylenediamine (TEMED) was purchased from ThermoScientific. Ammonium persulfate (APS) was purchased from Aqua Solutions. Polyethylene(glycol) diacrylate (PEGDA; Mw = 400) was purchased from PolySciences Inc. 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) was purchased from Advanced Chemtec. Hydrochloric acid (37.5%) and glycine were purchased from Fisher Scientific. Glass microscope slides (3” × 1” × 1.2 mm) were purchased from Fisherbrand. Ethanol (EtOH) was purchased from Decon Laboratories Inc. Water (dd-H2O) was deionized using Milli-Q Advantage A-10 water purification system (Millipore, U.S.A.). When required, degassing was performed by sparging with a continuous argon stream for 15 min for small volumes and 2 h for large volumes. Coverglass slips (Thickness #1, Diameter 25mm) were purchased from Electron Microscopy Sciences. Rain-X hydrophobic spray manufactured by ITW (Global Bands, TX) was purchased from Home Depot.

Parameswar A.V., Dikshit K.V., Movafaghi S., Bruns C.J, & Goodwin A.P. (2020). Mechanochemistry Activated Covalent Conjugation Reactions in Soft Hydrogels Induced by Interfacial Failure. ACS applied materials & interfaces, 13(1), 1486-1492.

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Osthole Modulates the IKK/NF-κB Pathway

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Osthole was obtained from Aladdin (Aladdin, Shanghai, China). Sodium dodecyl sulfate (SDS), ammonium persulfate (APS), TritonX-100, Tris base, glycine, hematoxylin and eosin were purchased from Sigma Chemical Co (St Louis, MO, USA). Bovine type II collagen was bought from Chondrex (Chondrex, Redmond, WA). Tetramethylethylenediamine (TEMED) and Tween 20 were purchased from Thermo Fisher Scientific (Waltham, MA). 30% Acrylamide/Bis Solution (37.5:1) and loading buffer were obtained from Bio-Rad Laboratories (Richmond, Calif., USA). Pre-stained protein marker was obtained from Fermentas (Glen Burnie, MD, USA). PVDF membrane was purchased from Millipore (Bedford, MA, USA). Antibodies to IKKα (11930), IKKβ (#8943), p-IKKα/β (#2697), IκBα(#4814), p-IκBα(#2859), p65 (#8242), p-p65 (#3033), SAPK/JNK (#9295), p-SAPK/JNK (#4668), Erk1/2 (#4695), p-Erk1/2 (#4370), p38 (#8690), p-p38 (#4511), β-actin (#5174), goat anti-rabbit horseradish peroxidase (HRP) (#7074) and RIPA cell lysis buffer (#9806) were purchased from CST (Boston, MA).

Xu R., Liu Z., Hou J., Huang T, & Yang M. (2018). Osthole improves collagen-induced arthritis in a rat model through inhibiting inflammation and cellular stress. Cellular & Molecular Biology Letters, 23, 19.

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Ctr1 Cu(I) Transporter Peptide Characterization

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The C-terminal 13 amino acids of the human Ctr1 Cu(I)-transporter (Ctr1c), KKAVVVDITEHCH, were custom-ordered as a lyophilized peptide from Sigma. DTT (dithiotreitol), TCEP-HCl (tris(2-carboxyethyl)phosphine), and IPTG (isopropyl 1-thio-β-d-galactopyranoside) were purchased from GoldBio. Tris-base, mono- and dibasic sodium phosphate, sodium acetate, ammonium persulfate (APS), EDTA, β-mercaptoethanol (BME), and sodium chloride were acquired from Fisher. Nitroblue terazolium and bathocuproinedisulfonic acid (BCS) were purchased from Alfa Aesar. Primers for site-directed mutagenesis and zinc sulfate heptahydrate were bought from Sigma. Maleimide-polyethyleneglycol 2000 was purchased from nanocs. Tetramethylethylenediamine (TEMED) was purchased from Thermo Scientific, while riboflavin was acquired from Acros Organics. Alexa-488-succinimidyl ester was purchased from Life Technologies. Cu(I)-(CH₃CN₄)PF₆ was purchased from Strem Chemicals. His-Trap Nickel affinity columns, SQ (anion exchange) columns, and gel filtration columns were purchased from GE LifeSciences.

Skopp A., Boyd S.D., Ullrich M.S., Liu L, & Winkler D.D. (2019). Copper–zinc superoxide dismutase (Sod1) activation terminates interaction between its copper chaperone (Ccs) and the cytosolic metal-binding domain of the copper importer Ctr1. Biometals, 32(4), 695-705.

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