Poly(hydroxybuyrate) (PHB) was purchased from Goodfellow (Huntington, England). Fiber-grade polylactic acid (PLA) (NatureWorks 6202D) was purchased from NatureWorks (Minnetonka, MN). Glutathione was purchased from GoldBio (St. Louis, MO). Acetone was purchased from VWR (Radnor, PA). Ethylenediaminetetraacetic acid (EDTA), Dulbecco’s modified Eagle’s medium (DMEM), sodium nitrite, DAPI, and the CCK-8 kit were purchased from Sigma Aldrich (St. Louis, MO). Penicillin-Streptomycin and fetal bovine serum were purchased from Gibco-Life Technologies (Grand Island, NY). HCl was purchased from Fisher Scientific (Hampton, NH). Hexafluoro-2-propanol (HFIP) was purchased from Oakwood Scientific (Estill, SC). Phosphate buffered saline (PBS), pH 7.4, composed of 138 mM NaCl, 2.7 mM KCl, and 10 mM sodium phosphate, was used for all in vitro experiments. The lactic dehydrogenase (LDH) kit was purchased from Roche Life Sciences (Indianapolis, IN). The bacteria strain S. aureus (ATCC 5538) and NIH/3T3 mouse fibroblasts (ATCC 1658) were received from American Type Culture Collection (Manassas, VA). Alexa Fluor 488 phalloidin was purchased from Thermo Fisher Scientific (Waltham, MA).
Glutathione (gsh)
Manufactured by GoldBio
Sourced in United States
Glutathione is a tripeptide composed of the amino acids cysteine, glutamic acid, and glycine. It is a naturally occurring antioxidant found in living organisms that plays a crucial role in cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Tris hcl ph 8, by Thermo Fisher Scientific (2 mentions)
Edta ph 8, by Merck Group (2 mentions)
Glutathione reduced, by GoldBio (2 mentions)
L arginine, by Merck Group (2 mentions)
Acetone, by Avantor (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Lactic dehydrogenase ldh kit, by Roche (1 mentions)
Cck 8 kit, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Nih 3t3 mouse fibroblast, by American Type Culture Collection (1 mentions)
S aureus atcc 5538, by American Type Culture Collection (1 mentions)
Hitrap q column, by GE Healthcare (1 mentions)
Trioctyl trimellitate, by Merck Group (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Nitrogen and oxygen gas cylinders, by Airgas (1 mentions)
Milli q filter, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
4 protocols using glutathione (gsh)
1
Synthesis and Characterization of PHB-PLA Biomaterial
2
Recombinant pHLA Complex Production
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DNA plasmids encoding HLA‐A*03:01 α‐chain and β2‐microglobulin were transformed separately into a BL21 strain of Escherichia Coli, as previously described.63 Recombinant proteins were expressed individually, where inclusion bodies were extracted and purified from the transformed E. coli cells. Soluble pHLA complexes were produced by refolding 30 mg of HLA‐A*03:01 with 10 mg of β‐2‐microglobulin and 5 mg of either NP265‐IAV, NP323‐IBV or NP270‐ICV peptide (Genscript, Piscataway, USA) into a buffer of 3 m Urea (Univar solutions, USA), 0.5 m L‐Arginine (Sigma‐Aldrich), 0.1 m Tris–HCl pH 8.0 (Fisher Bioreagents), 2.5 mm EDTA pH 8.0 (Sigma‐Aldrich, St Louis, USA), 5 mm Glutathione (reduced) (Goldbio, St Louis, USA) and 1.25 mm Glutathione (oxidised; Goldbio) for 3 h. The refold mixture was dialysed into 10 mm Tris–HCl pH 8.0 (Fisher Bioreagents), and soluble pHLA was purified using anion exchange chromatography using a HiTrapQ column (GE Healthcare).
3
Recombinant Peptide-HLA-B*15:01 Complex Production
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pET-30a(+) DNA plasmids encoding HLA-B*15:01 α-chain and human β2-microglobulin were transformed separately into BL21 E. coli and expressed. Inclusion bodies containing the individually expressed recombinant proteins were extracted, purified, and quantified as previously described.35 (link) Soluble peptide-HLA-B*15:01 complexes were produced by refolding 30 mg of HLA-B*15:01 α-chain with 10 mg of β2-microglobulin and 5 mg of peptide (Genscript, Piscataway, USA) into a buffer of 0.5 M L-Arginine (Sigma-Aldrich, St Louis, USA), 0.1 M Tris-HCl pH 8.0 (Fisher Bioreagents, Waltham, USA), 2.5 mM EDTA pH 8.0 (Sigma-Aldrich, St Louis, USA), 5 mM Glutathione (reduced) (Goldbio, St Louis, USA), 1.25 mM Glutathione (Goldbio, St Louis, USA). The peptides selected are summarized in Table 1 . The refold mixture was dialyzed at 4°C in 10 mM Tris-HCl pH 8.0 (3 times for 12 hours) and soluble pHLA complexes were purified via anion exchange chromatography.
4
Synthesis and Characterization of Biocompatible Materials
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