For direct tissue matrix-assisted laser desorption/ionization-Fourier transform mass spectrometry (MALDI-FTMS), midguts were removed from the thorax of cold-anesthetized crabs and pinned in a Sylgard-lined Petri dish containing physiological saline; small pieces of anterior midgut caecum (AMC) and posterior midgut caecum (PMC) were isolated via manual microdissection. Tissue fragments were rinsed sequentially in two 12 μL droplets of 0.75 M fructose (Sigma-Aldrich, St. Louis, MO, USA; 99%), placed on one face of a ten-faceted probe tip, and then sliced 10–20 times with a 0.1 mm needle. The macerated tissue was then gathered together and covered with a 0.5 μL droplet of 1.0 M 2,5-dihydroxybenzoic acid (DHB; Sigma-Aldrich; 98%, sublimed prior to use), prepared in 1:1 acetonitrile (Fisher Scientific, Pittsburg, PA, USA; HPLC grade) and water containing 2% (v/v) phosphoric acid.
2 5 dihydroxybenzoic acid dhb
Manufactured by Merck Group
Sourced in United States, Germany, Poland, Belgium, United Kingdom, France, Switzerland
2,5-dihydroxybenzoic acid (DHB) is a chemical compound that is commonly used as a matrix for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. It is a crystalline solid with the molecular formula C₇H₆O₃.
Automatically generated - may contain errors
Lab products found in correlation
Trifluoroacetic acid tfa, by Merck Group (7 mentions)
9 aminoacridine 9 aa, by Merck Group (4 mentions)
Ethanol, by Thermo Fisher Scientific (4 mentions)
Acetonitrile, by Thermo Fisher Scientific (3 mentions)
Acetonitrile acn, by Merck Group (3 mentions)
N n dimethylaniline dma, by Merck Group (3 mentions)
Methanol, by Merck Group (3 mentions)
Chloroform, by Merck Group (3 mentions)
Trifluoroacetic acid, by Avantor (2 mentions)
Acetic anhydride, by Merck Group (2 mentions)
Q exactive plus orbitrap, by Thermo Fisher Scientific (2 mentions)
Poly l lysine coated, by Merck Group (2 mentions)
Indium tin oxide conductive slides, by Bruker (2 mentions)
349 nm laser, by Spectra-Physics (2 mentions)
Hplc grade ethanol, by Merck Group (2 mentions)
Fetuin from fetal bovine serum, by Merck Group (2 mentions)
Iodoacetamide, by Merck Group (2 mentions)
Milli q system, by Merck Group (2 mentions)
Ammonium acetate, by Merck Group (2 mentions)
α cyano 4 hydroxycinnamic acid α chca, by Merck Group (2 mentions)
69 protocols using 2 5 dihydroxybenzoic acid dhb
1
MALDI-FTMS Analysis of Crustacean Midgut Tissue
2
Extraction and Analysis of Natural Dyes
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Indigo and alizarin (97%) were purchased from ACROS Organics; indirubin (≥98%), 9-aminoacridine (9-AA, ≥99.5%), erythrosin B (≥95%), 2-iodobenzoic acid (≥99%) and Universal MALDI matrix [1:1 mixture of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxy-cinnamic acid (α-CCA)] were obtained from Sigma-Aldrich, carminic acid (≥96%) from Fluka Analytical, lucidin from Cfm Oskar Tropitzsch GmbH (Marktredwitz), tetrahydrofuran (HPLC grade) from Roth, formic acid (99–100%), trifluoroacetic acid, ethanol, methanol and acetonitrile (all solvents HPLC gradient grade) from VWR, and TLC (thin-layer chromatography) plates (ALUGRAM Sil G) were purchased from Macherey-Nagel. Purpurin has been obtained from Aldrich’s collection of rare chemicals and rubiadin has been synthesized according to procedures of Takano et al.33 (link). Double distilled water was produced in a distillation apparatus bought from Gebr. Rettberg GmbH. An extended tabby weave with blue warps (wool fibers previously vat dyed using natural indigo) and red wefts (wool fibers previously mordant dyed using alum and cochineal) was handcrafted by a textile archaeologist. Recent animal- and plant-derived fibers dyed with synthetic indigo or E120, a food dye extract from scale insects (Dactylopius coccus Costa), were used for first tests (dyeing processes were performed as previously reported28 (link),34 (link)).
3
Synthesis of Functionalized Peptides
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9-fluorenylmethoxy-carbonyl (Fmoc)-protected amino-acids were from GenScript USA Inc. 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronoium hexafluorphosphate (HBTU), [N1-(Fmoc)-1,13-diamino-4,7,10-trioxatridecan-succinamic acid (Fmoc-TTDS) and Fmoc-Lys(N3)-OH were from Iris Biotech GmbH. 1-hydroxy-benzotriazole (HOBt), N,N′-diisopropylethylamine (DIPEA), N-methylpiperidine, acetic anhydride, pyridine, biotin, 4-pentynoic acid, Triisopropylsilane (TIPS), TRIZMA base and 2,5-Dihydroxybenzoic acid (DHB) were from Sigma-Aldrich. Trifluoroacetic acid (TFA), acetonitrile (MeCN), dichloromethane (DCM), N,N-dimethylformamide (DMF) and diethyl ether were from Fisher scientific. Fmoc-Dab(Alloc)-OH was from Bachem AG. N-methyl-2-pyrrolidone (NMP) was from Applied Biosystems.
4
Phospholipid Standards for Biochemical Analysis
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Phospholipid standards: 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine, 1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt), 1,2-dimyristoyl-glycero-3-phosphocholineand cardiolipin solution from a bovine heart were purchased from Avanti® Polar Lipids, Inc. (Alabaster, AL, USA) or Sigma-Aldrich.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2,5-dihydroxybenzoic acid (DHB) were purchased from Sigma-Aldrich, tryptone soya broth (TSB) was from BTL (Lodz, Poland). Surfactin and iturin A were obtained from Sigma-Aldrich. The other chemicals came from J.T. Baker, Fluka and POCh (Gliwice, Poland). All the chemicals were high purity grade reagents.
5
MALDI-MS Imaging on Tissue Sections
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Tissue sections of 12 μm thickness were thaw-mounted onto poly-l -lysine-coated (Sigma-Aldrich, Saint Louis, MO, USA) indium-tin oxide (ITO) conductive slides (Bruker Daltonics, Billerica, MA, USA) and stored at − 80 °C until use. ITO slides were thawed under vacuum for 15 min, and 8 layers (2 layers at 5 µl/min followed by 6 layers at 10 µl/min) of 2,5-dihydroxybenzoic acid (DHB (Sigma-Aldrich, Saint Louis, MO, USA) 30 mg/ml in MeOH:H2O 70:30, 0.2% TFA) were applied using a SunCollect (SunChrom, Friedrichsdorf, Germany) automated matrix spraying system (4 bar, 50 mm z axis height). The tissue sections were then dried under vacuum for 15 min prior to MALDI MSI data acquisition. MALDI MSI was performed with an EP-MALDI source [27 (link)] (Spectroglyph, LLC., Kennewick, WA, USA) equipped with a 349-nm laser (Spectra-Physics, Santa Clara, CA, USA), coupled with an Orbitrap QExactive Plus (Thermo Fisher Scientific). The laser was operated at 1.65 A and 500 Hz, the ion source pressure was 7.2 Torr, and the MSI pixel size was 35 × 35 µm. Mass spectra were acquired in the range 150–2000 m/z at 70,000 resolving power. The position file was aligned to the raw file using Image Insight (v. 0.1.0.11550, Spectroglyph, LLC).
6
MALDI Imaging of Metabolites in SVZ
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The tissue blocks were warmed to −20 °C, mounted onto a chuck using Optimal Cutting Temperature (OCT; Sakura Finetek, Torrance, CA), sectioned at 12 µm thickness on a Bright OTF5000 Cryostat (A-M Systems, USA) at −20°C and thaw-mounted onto pre-cooled indium tin oxide-coated MALDI glass slides (Hudson Surface Technology, USA). All sections were obtained from the central region of the rostral-caudal axis of the SVZ. The sections were dehydrated in a vacuum desiccator (Jencons, USA) for 1 hour and then rinsed with ammonium formate (50 mM) as described60 (link) to reduce sodium- and potassium-adducted species and to increase signal intensity and the signal-to-noise ratio. For MALDI imaging in positive mode, 2,5-dihydroxybenzoic acid (DHB, Sigma-Aldrich, St Louis, MO) was deposited onto the samples using an in-house vacuum sublimation apparatus for 10 min at approximately 50 mTorr and 110 °C. For MALDI imaging in negative mode, 1,5-diaminonaphthalene (DAN, Sigma-Aldrich, St Louis, MO) was similarly deposited for 5 min at approximately 50 mTorr and 140 °C.
7
Synthesis of Metal Nanoparticles and Biomolecules
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Ammonium hydroxide (28–30%), tetraethyl orthosilicate (TEOS, 96%), ethanol absolute (99.7%), silver nitrate (99.5%), tetrachloroauric acid tetrahydrate (HAuCl4·4H2O, 47.8%), hydrochloric acid (HCl, ≈36–38%), ethanol absolute (99.7%), sodium chloride (99.5%), FeCl3•6H2O (>97%), trisodium citrate (>99%), sodium acetate (>99%), methanol (99.8%), acetonitrile (99.5%), and ethylene glycol were purchased from Sinopharm Chemical Reagent Beijing Co., Ltd (Beijing, China). Polyvinylpyrrolidone (PVP, MW 40 kDa), phenylalanine (Phe, 98%), BSA, Pluronic F127, 2,5‐dihydroxybenzoic acid (DHB), alpha‐cyano‐4‐hydroxycinnamic acid (CHCA), IA (98%), and cholest‐5‐ene were ordered from Sigma‐Aldrich (St. Louis, MO, USA). Phenylacetaldehyde (Phen, 95%), OA (99%), and ascorbic acid (99%) were ordered from Aladdin Reagent Co., Ltd (Shanghai, China). AA (97%) was bought from Shanghai Macklin Biochemical Co., Ltd. Heptacosanoic acid (98%) was purchased from TCI Development Co., Ltd (Shanghai, China). [2H5]‐Phe and [2H5]‐Tyr was bought from Cambridge Isotope Labs (America). All aqueous solutions throughout the experiments were prepared with deionized water (18.2 MΩ·cm, Milli‐Q, Millipore, GmbH).
8
Glycoprotein Analysis via Mass Spectrometry
9
Human IgG N-Glycan Analysis
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Human IgG
from serum, bovine
fetuin, 2-aminobenzamide (2-AB), 2-aminobenzoic acid (2-AA), 2-13[C6]-aminobenzoic acid, iodoacetamide, dithiothreitol,
dimethyl sulfoxide (DMSO), methylamine hydrochloride, (7-azabenzotriazol-1-yloxy)trispyrrolidinophosphonium
hexafluorophosphate (PyAOP), 4-methylmorpholine, sodium hydroxide,
cellulose (medium fibrous), iodomethane, and 2,5-dihydroxybenzoic
acid (DHB) were purchased from Sigma (St. Louis, MO). N-Glycosidase
F (PNGase F) was obtained from New England Biolabs (Ipswich, MA).
The Viva Spin series of spin filters (10K and 30K MWCO) were purchased
from Sartorius Stedium Biotech (Aubagne, France).
from serum, bovine
fetuin, 2-aminobenzamide (2-AB), 2-aminobenzoic acid (2-AA), 2-13[C6]-aminobenzoic acid, iodoacetamide, dithiothreitol,
dimethyl sulfoxide (DMSO), methylamine hydrochloride, (7-azabenzotriazol-1-yloxy)trispyrrolidinophosphonium
hexafluorophosphate (PyAOP), 4-methylmorpholine, sodium hydroxide,
cellulose (medium fibrous), iodomethane, and 2,5-dihydroxybenzoic
acid (DHB) were purchased from Sigma (St. Louis, MO). N-Glycosidase
F (PNGase F) was obtained from New England Biolabs (Ipswich, MA).
The Viva Spin series of spin filters (10K and 30K MWCO) were purchased
from Sartorius Stedium Biotech (Aubagne, France).
10
Glucose Detection via CeO2 Nanoparticles
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All DNA oligonucleotides used in this study were synthesized and purified with high performance liquid chromatography (HPLC) by Bioneer® (Daejeon, Korea). The sequence information of oligonucleotides employed in this study is in Table S1 . The personal glucose meter (PGM) whose dynamic range for glucose detection is from 0.6 to 33 mM was purchased from Accu-Chek (Roche, Basel, Switzerland). Cerium (IV) oxide nanoparticle (CeO2 NP), sodium acetate, glucose, ethidium bromide (EtBr), hydroethidine, and 2,5-dihydroxybenzoic acid (DHB) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The deoxynucleoside triphosphate (dNTP) and i-TaqTM DNA polymerase were purchased from Intron Biotechnology Inc. (Daejeon, Korea). Ultrapure DNase/RNase-free distilled water (DW) purchased from Bioneer® was used in all experiments. All chemicals used in this study were of analytical grade.
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