For the preparation of the 5% and 20% EMP hydrogels, the monomer solvent—acryl-EMP (2.83 µmol, 5% or 11.32 µmol, 20%, respectively) with acrylamide (1.07 µmol, 7.6%, Nacalai Tesque, Kyoto, Japan) and N,N′-methylene bisacrylamide (25.9 µmol, 0.4%, Nacalai Tesque, Kyoto, Japan)—in 1 mL of 375 mM Tris-HCl pH 8.8 buffer were polymerized by adding ammonium persulfate (0.1%, Nacalai Tesque, Kyoto, Japan) and TEMED (0.02%, Kanto Chemicals, Tokyo, Japan). After adding ammonium persulfate and TEMED, the monomer solvent was poured in thin cassettes (1 mm thickness and 90 mm width) and polymerized at 20 °C. Non-crosslinked EMP hydrogels (NCE hydrogels) (5% and 20%) were prepared by using EMP instead of acryl-EMP. For EMP-NCE bilayer hydrogel preparation, the same volume of acryl-EMP monomer solvent was polymerized at 20 °C on top of the NCE hydrogel in the same thin cassettes.
N n methylene bisacrylamide
Manufactured by Nacalai Tesque
Sourced in Japan
N,N′-methylene bisacrylamide is a chemical compound used as a cross-linking agent in various applications. It is a white, crystalline solid with the chemical formula C7H10N2O2.
Automatically generated - may contain errors
Lab products found in correlation
Ammonium persulfate, by Nacalai Tesque (1 mentions)
Acrylamide, by Nacalai Tesque (1 mentions)
Guanidine hydrochloride, by Nacalai Tesque (1 mentions)
Ultrapure tris, by Thermo Fisher Scientific (1 mentions)
Sodium acrylate, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Nacalai Tesque (1 mentions)
Triton x 100, by Nacalai Tesque (1 mentions)
Acrylamide aa, by Merck Group (1 mentions)
Proteinase k, by Nacalai Tesque (1 mentions)
Seaplaque agarose, by Lonza (1 mentions)
2 protocols using n n methylene bisacrylamide
1
Synthesis of EMP Hydrogels
2
Hydrogel-Based Cell Encapsulation Protocol
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Antibodies used in this study are listed in Supplemental Table S1. The following reagents were purchased from the manufacturers as noted: sodium acrylate (SA) (#408220) from Sigma-Aldrich; acrylamide (AA) (#00809-85), N,N′-methylenebisacrylamide (#22402-02), ammonium persulfate (APS) (#02627-34), N,N,N′,N′-tetramethylethylenediamine (TEMED) (#33401-72), guanidine hydrochloride (#17318-95), EDTA (#15105-35), Triton X-100 (#12967-45), and proteinase K (#29442-14) from Nacalai Tesque; 6-((acryloyl)amino)hexanoic acid, succinimidyl ester (Acryloyl-X, SE) (#A20770), and UltraPure Tris (#15504-020) from Invitrogen; and SeaPlaque Agarose (#50100) from Lonza.
The monomer solution containing 8.6 g/100 ml of SA, 2.5 g/100 ml of AA, 0.15 g/100 ml of N,N′-methylenebisacrylamide, 11.7 g/100 ml of sodium chloride, and 1× phosphate-buffered saline (PBS) was prepared and stored at 4°C until use. Digestion buffer containing 50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.5% Triton X-100, and 0.8 M guanidine hydrochloride was prepared and stored at 4°C until use.
The following materials were purchased from the manufacturers: Grace Bio-Labs CultureWell removable chambered cover glass (Sigma-Aldrich; #GBL112358), six-well glass-bottom plates with high-performance #1.5 cover glass (Cellvis; P06-1.5H-N), and coverslips (9-mm diameter) (Matsunami Glass; #CS01029).
The monomer solution containing 8.6 g/100 ml of SA, 2.5 g/100 ml of AA, 0.15 g/100 ml of N,N′-methylenebisacrylamide, 11.7 g/100 ml of sodium chloride, and 1× phosphate-buffered saline (PBS) was prepared and stored at 4°C until use. Digestion buffer containing 50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.5% Triton X-100, and 0.8 M guanidine hydrochloride was prepared and stored at 4°C until use.
The following materials were purchased from the manufacturers: Grace Bio-Labs CultureWell removable chambered cover glass (Sigma-Aldrich; #GBL112358), six-well glass-bottom plates with high-performance #1.5 cover glass (Cellvis; P06-1.5H-N), and coverslips (9-mm diameter) (Matsunami Glass; #CS01029).
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