Recombinant human insulin was purchased from Dongbao Enterprise Group Co., Ltd. (Tonghua, Jilin, China). Lecithin (70% phosphatidylcholine/30% phosphatidyl-ethanolamine, lipoid phospholipid) was obtained from Shanghai Toshisun Biology and Technology Co., Ltd. (Shanghai, China). Sodium deoxycholate and D-(+)-trehalose dehydrate purchased from Sigma (St. Louis, MO, USA), α-lactose monohydrate purchased from Aladdin (Shanghai, China). Other chemicals and solvents were of analytical or chromatography grade.
D trehalose dehydrate
Manufactured by Merck Group
Sourced in United States
D-(+)-trehalose dehydrate is a chemical compound that is a disaccharide sugar. It is a structural isomer of sucrose, with the two glucose molecules connected in an alpha,alpha-1,1-linkage. D-(+)-trehalose dehydrate is used as a laboratory reagent and in various industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium azide, by Merck Group (6 mentions)
Curcumin, by Merck Group (2 mentions)
Ultrapure water, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Betaine, by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Sodium deoxycholate, by Merck Group (1 mentions)
Magnesium chloride hexahydrate, by Junsei (1 mentions)
3 aminopropyltriethoxysilane, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Klenow fragment exo, by New England Biolabs (1 mentions)
Polyethylene glycol, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Purelab flex 4, by Elga LabWater (1 mentions)
1 2 dipalmitoyl sn glycero 3 phosphocholine, by Lipoid (1 mentions)
Hepes, by Merck Group (1 mentions)
Cholesterol, by Merck Group (1 mentions)
Cytochrome c, by Abcam (1 mentions)
Calnexin, by Santa Cruz Biotechnology (1 mentions)
17 protocols using d trehalose dehydrate
1
Insulin Formulation Protocol
2
Biochemical Reagents Procurement Protocol
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BSA Fraction V was purchased from Biosesang (Kyung-gi, Korea). Sucrose, d -(+)-trehalose dehydrate, and 3-aminopropyl-triethoxysilane were purchased from Sigma-Aldrich (St Louis, MO, USA). Magnesium chloride hexahydrate and sodium phosphate dibasic anhydrate were obtained from Junsei Chemical Co., Ltd. (Tokyo, Japan). Ethanol was purchased from Merck KGaA (Darmstadt, Germany). All other chemicals were of analytical grade, and all solvents were of HPLC grade.
3
Massively Parallel DNA Sequencing Array
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All possible 65,536 8-base-pair (bp) DNA sequences were assembled into a maximally compact de Bruijn sequence that was subsequently divided over 1457 oligonucleotides. Each 70-bp-long oligonucleotide contained 45 overlapping 8-mers, a 3-bp GC clamp at the 5′ end, and an identical 14-bp sequence at the 3′ end for Cy5 labeling and primer extension (Fordyce et al. 2010 (link)). Sequences were hybridized to a Cy5-labeled oligonucleotide and extended using a Klenow fragment (exo−) (New England Biolabs) to produce Cy5-labeled dsDNA (Fordyce et al. 2010 (link)). Cy5-labeled dsDNA was diluted to a final concentration of 1.25 μM. Each sample solution contained 0.125% of poly(ethylene glycol) (Aldrich) and D-trehalose dehydrate (Sigma) at 12.5 mg/mL in dH2O to prevent irreversible binding of the DNA to the glass as well as for visualization during alignment of the device to the DNA array. The oligos were spotted onto epoxy-coated glass substrates (CEL Associates) with a MicroGrid 610 (Bio Robotics) microarrayer using SMT-S75 silicone pins (Parallel Synthesis). Column and row pitch corresponded to the specific device. The device that we used contained 65 columns and 64 rows with a pitch of 281.25 µm by 562.5 µm, respectively. Finally, the arrays were aligned to the microfluidic device by hand under a stereoscope and bonded overnight on a heated plate at 80°C.
4
Microneedle Vaccine Patch Development
5
Liposome Formulation with Tetraether Lipids
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Curcumin (≥80%), cholesterol (≥99%), HEPES (≥99.5%), ethanol (absolute, ≥99.8%), D-(+)-trehalose dehydrate (≥99%) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich Chemie GmbH (Taufkirchen, Germany). The lipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were a gift from Lipoid GmbH (Ludwigshafen am Rhein, Germany). Fractions containing the polar tetraether lipids caldarchaeol (GDGT, approx. 10% of the extract) and calditoglycerocaldarchaeol (GDNT, approximately 90% of the extract) were extracted from the biomass of Sulfolobus acidocaldarius (Transmit GmbH, Gießen, Germany) [31 (link)]. The ends of GDNT contain phosphatidylmyoinositol and β-glucose respectively while GDGT contains phosphatidylmyoinositol and β-D-galactosyl-D-glucose on either ends respectively [32 (link)]. Ultrapure water from PURELAB® flex 4 (ELGA LabWater, High Wycombe, UK) was used for all experiments in this study. All solvents used were of analytical or HPLC grade.
6
Metal-Conjugated Antibody Staining for MIBI
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Antibodies were first screened by IHC on FFPE brain sections to select for the best clones. These were then conjugated to isotopic metal reporters as described previously40 (link),44 (link). Following conjugation antibodies were diluted in Candor PBS Antibody Stabilization solution (Candor Bioscience). Antibodies were either stored at 4°C or lyophilized in 100 mM D-(+)-Trehalose dehydrate (Sigma-Aldrich) with ultrapure distilled H2O for storage at −20°C. Before staining, lyophilized antibodies were reconstituted in a buffer of Tris (Thermo Fisher Scientific), sodium azide (Sigma-Aldrich), ultrapure water (Thermo Fisher Scientific) and antibody stabilizer (Candor Bioscience) to a concentration of 0.05 mg ml−1. The antibodies, metal reporters and staining concentrations are listed in Extended Data Table S1 . For detailed metal-antibody protocol MIBItag see dx.doi.org/10.17504/protocols.io.bhyej7te .
7
Trehalose Dehydrate-Induced Autophagy Regulation
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D-(+)-Trehalose dehydrate (purity >99 %), toluidine blue, tert-Butyl hydroperoxide solution (TBHP), bafilomycin A1 (Baf), chloroquine (CQ) and type II collagenases were purchased from Sigma-Aldrich (St Louis, MO, USA). The primary antibody against p62, Tom20, BNIP3, PGAM5, Drp1, SOD2, PARP, cytochrome C, caspase 9, 8-0HdG and β-actin were from Abcam (Cambridge, UK), LC-3 antibody was from Sigma-Aldrich (St Louis, MO, USA). GRP78 and Calnexin antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); Anti-CHOP, goat anti-rabbit, and anti-mouse IgG-HRP were from Bioworld (OH, USA) and antibodies against Atg3, Atg7, Atg12, Beclin1, p-AKT, AKT, p-mTOR, mTOR, p-p70S6K, p70S6K, p-AMPK, AMPK, p-ULK1, ULK1, ubquintin, caspase 12, Bcl-2, Bax and cleaved-caspase3 were from Cell Signaling (Danvers, MA, USA); Alexa Fluor488 labeled and Alexa Fluor594 labeled Goat Anti-Rabbit IgG (H+L) second antibody were from Jackson ImmunoResearch (West Grove, PA, USA). 4′,6-diamidino-2-phenylindole (DAPI) was from Beyotime (Shanghai, China).
8
Antibody Conjugation and Storage Protocol
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Antibodies were conjugated to isotopic metal reporters as described previously18 (link). Following conjugation antibodies were diluted in Candor PBS Antibody Stabilization solution (Candor Bioscience). Antibodies were either stored at 4 °C or lyophilized in 100 mM D-( + )-Trehalose dehydrate (Sigma-Aldrich) with ultrapure distilled H2O for storage at −20 °C. Before staining, lyophilized antibodies were reconstituted in a buffer of Tris (Thermo Fisher Scientific), sodium azide (Sigma-Aldrich), ultrapure water (Thermo Fisher Scientific) and antibody stabilizer (Candor Bioscience) to a concentration of 0.05 mg ml−1. The antibodies, metal reporters and staining concentrations are listed in Extended Data Table 2 . A limitation of this study is that we did not have an antibody for labeling bacteria due to the inherent difficulty of antibody-based detection of Mtb in FFPE tissue.
9
Trehalose Coating on Copper Substrate
10
Curcumin-loaded PLGA Nanoparticles
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Poly(Lactide-co-Glycolide) acid (PLGA) with a 50:50 copolymer ratio and free ester end groups (Resomer® RG 504, MW 38,000–54,000), Poly(Vinyl) Alcohol (PVA, Mowiol® 4–88 MW 31,000), D-mannitol, D-(+)-trehalose dehydrate, curcumin and ethyl acetate were obtained from Sigma Aldrich and used without further purification.
Alexa Fluor® 488 goat anti-mouse IgG1 (isoform γ1, IgG1-488) was purchased from Life Technologies and diluted up to 0.3 mg/mL in a medium containing 0.1 M sodium phosphate, 0.1 M NaCl and 5.0 mM sodium azide (Sigma Aldrich). Bidistilled water (MilliQ, Millipore) was used in all the experiments.
Alexa Fluor® 488 goat anti-mouse IgG1 (isoform γ1, IgG1-488) was purchased from Life Technologies and diluted up to 0.3 mg/mL in a medium containing 0.1 M sodium phosphate, 0.1 M NaCl and 5.0 mM sodium azide (Sigma Aldrich). Bidistilled water (MilliQ, Millipore) was used in all the experiments.
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