Lipids used in this study that were purchased from Avanti Polar lipids (Alabaster, AL) were as follows: DPPE (16:0, Powder), DOPC (18:1, Chloroform), BSM (predominant 18:0, Porcine, Chloroform) and CHOL (ovine wool, ≥98%). For the fluorescence imaging, Texas Red 1,2-Dihexadecanoyl-sn-Glycero-3-Phosphoethanolamine, TR-DHPE was purchased from Invitrogen (Carlsbad, CA). DPPE was dissolved in a solvent, which is a 3:1 (vol/vol) mixture of chloroform (Sigma-Aldrich, CHROMASOLV Plus for HPLC, purity ≥99.9%) and methanol (Sigma-Aldrich, CHROMASOLV Plus for HPLC, purity ≥99.9%) at a final concentration of 1 mg ml−1. DOPC, BSM and CHOL were mixed in a 1:1:1 (mol/mol) solution at a final concentration of 1 mg ml−1 in chloroform. A trace amount (1 wt%) of TR-DHPE was added to the mixture for imaging purposes. All lipids were stored in a deep freezer (−50 °C) until use. Buffer salts were purchased from Sigma-Aldrich (St Louis, MO), mixed and dissolved in Milli-Q water (Millipore, Billerica, MA) at final concentrations of 100 mM Sodium nitrate (ReagentPlus, purity ≥99.0%), 10 mM Tris(hydroxymethyl)aminomethane (ACS reagent, purity ≥99.8%) and 2 mM Calcium nitrate tetrahydrate (purity ≥99.0%) at a pH of 7.5.
Chromasolv plus
Manufactured by Merck Group
Sourced in Germany, Italy
Chromasolv Plus is a high-purity solvent used in analytical and chromatographic applications. It is designed to provide consistent and reliable performance in various laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Milli q water, by Merck Group (3 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Ab26245, by Abcam (1 mentions)
Ab97043, by Abcam (1 mentions)
Oasis hlb spe cartridge, by Waters Corporation (1 mentions)
Hplc grade, by Merck Group (1 mentions)
0.45 μm syringe filter, by Cytiva (1 mentions)
1200 series, by Waters Corporation (1 mentions)
Ammonium hydroxide, by Thermo Fisher Scientific (1 mentions)
Tetraethyl orthosilicate, by Thermo Fisher Scientific (1 mentions)
2.2 bipyridine, by Thermo Fisher Scientific (1 mentions)
Texas red 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine tr dhpe, by Thermo Fisher Scientific (1 mentions)
Polydiallyldimethylammonium chloride, by Merck Group (1 mentions)
Luria bertani broth, by Merck Group (1 mentions)
Anhydrous toluene, by Merck Group (1 mentions)
Reagentplus, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
Emplura, by Merck Group (1 mentions)
Toluene, by Merck Group (1 mentions)
Tetrahydrofuran, by Merck Group (1 mentions)
21 protocols using chromasolv plus
1
Lipid Preparation for Fluorescence Imaging
2
Metabolome extraction from co-cultured S. costatum and K. algicida
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
S. costatum cultures reared in 12 C or 13 C medium were co-cultured in triplicates (185 mL each) with the K. algicida added to a final OD550 of 0.01. After 6 days the lysed cultures were gently filtered (0.2 µm pore size) and the flow-through extracted for metabolome analysis as follows.
Solid phase extraction cartridges (Chromabond easy, Macherey-Nagel, Düren, Germany) were equilibrated with 4 mL of methanol (Chromasolv © Plus, Sigma-Aldrich, Munich, Germany) and 4 mL of water (Chromasolv © Plus, Sigma-Aldrich, Munich, Germany) before the filtrate (170 mL) was applied using vacuum with a flow rate < 1 L h -1 . The cartridge was washed with 4 mL water, air-dried and then extracted via gravity flow using 2 mL of methanol followed by 2 mL of methanol/tetrahydrofuran 1:1 (tetrahydrofuran HiPerSolv, VWR, Dresden, Germany). This extract was frozen until further chemical analysis.
Solid phase extraction cartridges (Chromabond easy, Macherey-Nagel, Düren, Germany) were equilibrated with 4 mL of methanol (Chromasolv © Plus, Sigma-Aldrich, Munich, Germany) and 4 mL of water (Chromasolv © Plus, Sigma-Aldrich, Munich, Germany) before the filtrate (170 mL) was applied using vacuum with a flow rate < 1 L h -1 . The cartridge was washed with 4 mL water, air-dried and then extracted via gravity flow using 2 mL of methanol followed by 2 mL of methanol/tetrahydrofuran 1:1 (tetrahydrofuran HiPerSolv, VWR, Dresden, Germany). This extract was frozen until further chemical analysis.
3
Spectroscopic Characterization of HOCl-Modified Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unless otherwise stated, all chemicals were purchased from commercial sources and used as received. UPLC buffers were prepared using HPLC grade methanol (VWR) and tetrahydrofuran (Chromasolv Plus, Sigma Aldrich). Aqueous solutions were prepared using MilliQ water and buffers were prepared from 0.5 M sodium phosphate buffer at pH 7.4, or PBS, and chelex-treated prior to use to remove trace metal ions. Cl− and I− solutions were prepared using NaCl and KI (Sigma Aldrich). The concentration of stock H2O2 and HOCl solutions (Sigma Aldrich) were determined spectrophotometrically (H2O2 in H2O, ε240 43.6 M−1 cm−1; HOCl in 0.5 M NaOH, ε292 350 M−1 cm−1). Immunoblotting and ELISA were performed using the following monoclonal antibodies (mAb): anti-fibronectin A17 (ab26245, Abcam), anti-fibronectin A32 (CSI 005-32-02, Thermo Fisher Scientific), 2D10G9 (raised against HOCl-modified protein; a kind gift from A. Prof Ernst Malle, Medical University of Graz, Austria) and anti-dityrosine (Clone 1C3 monoclonal antibody, Nordic Biosite) [55 (link)]. Detection of immune complexes from immunoblotting was achieved with ECL anti-rabbit IgG horseradish peroxidase-linked whole antibody (NA934V, GE Healthcare UK). Detection for ELISA was achieved with a polyclonal rabbit anti-mouse IgG alkaline phosphate conjugated antibody (ab97043, Abcam).
4
Extraction and Analysis of Marine Exudates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 150 mL of filtered medium from each culture flask was transferred to sterile and cleaned 250 mL Erlenmeyer flasks, which were covered immediately with aluminum foil and cooled down to 4°C before solid phase extraction. Roseovarius sp. and Maribacter sp. exudates (n = 4, diluted to an equivalent OD600 = 0.05 with minimal medium) were prepared and stored in the same way. Before extraction, 15 nmol of caffeine dissolved in methanol [HPLC grade, Sigma–Aldrich, Chromasolv®Plus (≥99.9%)] was added to each sample as an internal standard. The medium was extracted on 60 mg Oasis® HLB-SPE cartridges (Waters, Eschborn, Germany), following the manufacturer’s instructions. Gentle vacuum was applied to the cartridges with a VisiprepTM SPE Vacuum Manifold (Sigma–Aldrich) to have a flow-through of ca. 1 drop per second. The cartridges were eluted three times with 1 mL of methanol. The 3 mL of eluate was stored in 4 mL vial glass at −80°C until further analysis. Medium blanks (n = 3) were prepare in the same way by extracting sterile F/2 medium. 1.5 mL of the eluate from each sample was transferred to a clean vial, evaporated under a stream of nitrogen, and dissolved in 50 μl of methanol. Two quality control (QC) samples were prepared by pooling 5 μl from each sample in one clean vial.
5
GC-MS Sample Preparation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Volumes equivalent to 5 × 106 cells (Orbitrap GC) or 4.2 × 107 cells (ISQ GC–MS) per sample were taken from each extract. Both cell numbers are below the recommended maximum cell number in our validated in-lab standard operating procedure (SOP) that has been published [51 (link)]. Pooled quality control (QC) samples were prepared by combining equal volumes from each extract [52 (link)]. All samples were dried under vacuum and reconstituted in 50 µL pyridine (CHROMASOLV Plus, Sigma-Aldrich, Munich, Germany) containing 20 mg/mL methoxyamine monohydrochloride (Sigma-Aldrich, Munich, Germany). Samples were heated to 60 °C for 1 h and stored at room temperature overnight. A volume of 50 µL of N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) (Thermo Scientific, Bremen, Germany) was added to each sample, and all samples were heated to 60 °C for 1 h. A C7–C40 alkane standard mix (Supelco, Munich, Germany) was added to one QC sample.
6
Molecular Weight Characterization of NHP407 and SHP407
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Number Average and Weight Average Molecular Weights ( and , respectively) and Polydispersity Index (D) of NHP407 and SHP407 samples were estimated using an Agilent Technologies 1200 Series (USA) instrument equipped with a Refractive Index detector (RID) and two Waters Styragel columns (HR2 and HR4). Analyses were conducted using tetrahydrofuran (THF, inhibitor-free, CHROMASOLV® Plus, for HPLC, ≥99.9%, Sigma Aldrich, Italy) as eluent at 35°C and 0.4 ml/min flow rate. SEC samples were prepared by filtering a polymer solution (2 mg/ml in THF) through a 0.45 μm syringe filter (Whatman). Registered RID signals as a function of elution time were then analyzed using Excel software (Microsoft Corporation) to estimate , , and D relative to a calibration curve based on polystyrene standards ( within the range 740–180,000 Da).
7
Synthesis of Organic Electrolytes for Energy Applications
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tetraethyl orthosilicate (TEOS, 98%) and ammonium hydroxide (28–30%) were purchased from Acros. C6F13COOH was purchased from P&M Invest and used without further purification. CH3CN (CHROMASOLV® Plus, ≥99.9%, by Acros Organics) was used without any preliminary purification. 2.2′-bipyridine (99%, from Acros Organic), and C6H6 (CHROMASOLV® Plus, ≥99.9%, by Sigma Aldrich) were used without any preliminary purification as well.
The ethanol and TEOS were purified by distillation.
Et4NBF4 was obtained by mixing an aqueous solution of Et4NOH (30–35%) with HBF4 for to a neutral indicator reaction. Et4NBF4 precipitated from the reaction mixture as white crystals, which were separated by filtering. The powder salt was further recrystallized from diethyl ether and dried for 2 to 3 days in a vacuum at 55 °C for dehydration.
The ethanol and TEOS were purified by distillation.
Et4NBF4 was obtained by mixing an aqueous solution of Et4NOH (30–35%) with HBF4 for to a neutral indicator reaction. Et4NBF4 precipitated from the reaction mixture as white crystals, which were separated by filtering. The powder salt was further recrystallized from diethyl ether and dried for 2 to 3 days in a vacuum at 55 °C for dehydration.
8
Adhesive Leaf Mounting for LAAPPI-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Milli-Q
water (Merck Millipore, Molsheim,
France) was used to adhere leaves onto microscope glass slides (see
below), whereas toluene (CHROMASOLV Plus, Sigma-Aldrich, Steinheim,
Germany) was used as a spray solvent in the negative ion LAAPPI-MS
measurements.
water (Merck Millipore, Molsheim,
France) was used to adhere leaves onto microscope glass slides (see
below), whereas toluene (CHROMASOLV Plus, Sigma-Aldrich, Steinheim,
Germany) was used as a spray solvent in the negative ion LAAPPI-MS
measurements.
9
Zinc Surface Modification Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Zinc sheets (1 mm thick, 99.99+%) were purchased from Goodfellow Ltd and cut into 2 × 1 cm2 pieces. Sodium bicarbonate (NaHCO3, purum p.a., 99.0%), 2-propanol (IPA, Chromasolv® Plus, for HPLC, 99.9%), hydrochloric acid (HCl, ACS reagent, 37%), benzyl-hexadecyl-dimetylammonium chloride (BAC, pure cationic surfactant), potassium bromide (KBr, FTIR grade, >99% trace metals basis), poly-diallyl-(dimethylammonium) chloride (PDDA, with an average molecular weight MW of 200,000–350,000, 20 wt.% in H2O), and Luria-Bertani (LB) broth (Miller, pH 6.8–7.2, 2.5% solution) were purchased from Merck-Sigma Aldrich (Milan, Italy). Aluminum oxide (Al2O3, purum p.a., 99.7%), for the mechanical polishing of zinc sheets, was obtained from Fluka Chemicals (Milan, Italy). Agar powder, meat extract, and peptone (from casein, pancreatic digest) were supplied by SIFIN Diagnostics Gmbh (Berlin, Germany). Milli-Q water was used in all experiments.
10
Matrimid 5218 Spin-Coating on Silicon Wafers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Matrimid 5218 US (Huntsman) (Figure 1 ), Cyclopentanone
(ReagentPlus, ≥99%,
Sigma-Aldrich), toluene (EMPLURA, >99%, Merck), n-hexane (EMPLURA, >95%, Merck), toluene anhydrous (99.8%, Sigma-Aldrich),
and n-hexane anhydrous (95%, Sigma-Aldrich) for the
BDS experiments were used as received.
Silicon wafers (100,
front side polished,
CZ test grade, Silchem) were used as a substrate for the spin-coated
layer. The wafers were stored under clean-room conditions until being
cut. Prior to use, the wafers were cleaned with acetone (Chromasolv
plus, for HPLC 99.9%, Sigma-Aldrich).
(ReagentPlus, ≥99%,
Sigma-Aldrich), toluene (EMPLURA, >99%, Merck), n-hexane (EMPLURA, >95%, Merck), toluene anhydrous (99.8%, Sigma-Aldrich),
and n-hexane anhydrous (95%, Sigma-Aldrich) for the
BDS experiments were used as received.
Silicon wafers (100,
front side polished,
CZ test grade, Silchem) were used as a substrate for the spin-coated
layer. The wafers were stored under clean-room conditions until being
cut. Prior to use, the wafers were cleaned with acetone (Chromasolv
plus, for HPLC 99.9%, Sigma-Aldrich).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!