H2228, AsPC-1, and A549 cells were cultured in 10-cm dishes under HSS or GG conditions at 37 °C for 3 days. Cells (1.0 × 106 cells) were washed twice with ice-cold 5% mannitol solution and covered with 1 mL of methanol containing 25 μM internal standards, methionine sulfate (FUJIFILM Wako Pure Chemical, Osaka, Japan), 2-(N-morpholino)-ethanesulfonic acid (FUJIFILM Wako Pure Chemical), and D-camphor-10-sulfonic acid (FUJIFILM Wako Pure Chemical). The collected samples (400 μL) were mixed with 400 μL of chloroform and 200 μL of Milli-Q water. The aqueous phase of the cell sample was then subjected to ultrafiltration. Metabolites of cells were quantified using CE-MS (Agilent Technologies, Santa Clara, CA, USA) as previously described with automatic integration software (MasterHands version 2.17.3.18, Keio University)37 (link) and analyzed using MetaboAnalyst 5.038 (link). A hierarchical clustering heatmap was generated using the Euclidean correlation. The fold change of each metabolite was measured by comparing the absolute value of change between the two groups. The small molecule pathway database (SMPDB) was used for metabolite set enrichment analysis (MSEA).
D camphor 10 sulfonic acid
Manufactured by Fujifilm
Sourced in Japan, United States
D-camphor-10-sulfonic acid is a chemical compound used in various laboratory applications. It is a white crystalline solid that is soluble in water and organic solvents. The compound has a specific function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
2 n morpholino ethanesulfonic acid, by Dojindo Laboratories (3 mentions)
3 aminopyrrolidine, by Merck Group (3 mentions)
Trimesate, by Fujifilm (2 mentions)
L methionine sulfone, by Fujifilm (2 mentions)
G6220a lc msd tof, by Agilent Technologies (2 mentions)
1100 series isocratic hplc pump, by Agilent Technologies (2 mentions)
Agilent g1607a ce esi ms sprayer kit, by Agilent Technologies (2 mentions)
Agilent g1603a ce ms adapter kit, by Agilent Technologies (2 mentions)
Ce ms, by Agilent Technologies (1 mentions)
2 n morpholino ethanesulfonic acid, by Fujifilm (1 mentions)
Agilent ce capillary electrophoresis system, by Agilent Technologies (1 mentions)
Ultrafreemc plhcc for metabolome analysis, by Human Metabolome Technologies (1 mentions)
Trimesic acid, by Fujifilm (1 mentions)
Agilent ce tofms system, by Agilent Technologies (1 mentions)
Methionine sulfone, by Fujifilm (1 mentions)
3 mm zirconia beads, by Biospec (1 mentions)
Methanol, by Fujifilm (1 mentions)
Acetonitrile, by Fujifilm (1 mentions)
Bde 209, by Merck Group (1 mentions)
N n diethyl 2 phenylacetamide, by Fujifilm (1 mentions)
7 protocols using d camphor 10 sulfonic acid
1
Metabolic Profiling of Cancer Cell Lines
2
Metabolite Extraction and Analysis of Melanoma Cells
3
Metabolome Analysis of Mrs2 Knockdown
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were transfected with a plasmid for Mrs2 knockdown by electroporation using Neon (Life Technologies). The cells were plated on a 100 mm dish, and grown for 3 days at 37 °C in a humidified atmosphere containing 5% CO2. Culture medium was changed every day. Procedure for sample preparation of metabolome analysis was previously described37 (link). Briefly, after washing cells twice with ice-cold 5% mannitol, metabolites were extracted by 1 mL ice-cold methanol containing internal standards (25 μM each of methionine sulfone (MetSul; Wako, Osaka, Japan), 2-(N-morpholino)ethanesulfonic acid (MES; wako), D-Camphor-10-sulfonic acid (CSA; Wako). 400 μL of collected extracts were transferred into another tube, mixed with 400 μL chloroform and 200 μL Milli-Q water, and centrifuged at 10,000 × g for 3 min at 4 °C. A 400 μL aliquot of the aqueous layer was centrifugally filtered through a 5 kDa cutoff membrane (UltrafreeMC-PLHCC for Metabolome Analysis; Human Metabolome Technologies, Yamagata, Japan) to remove proteins from samples, followed by the centrifugal-concentration at 42 °C. CE-MS experiments were performed using Agilent CE Capillary Electrophoresis System.
4
Quantitative Metabolite Analysis of Quadriceps
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A quantitative analysis of charged metabolites in the quadriceps femoris was performed using CE-TOFMS as described previously (66 (link), 67 (link)). Quadriceps femoris muscle samples (50 mg) were submerged in methanol (500 μl) containing internal standards [methionine sulfone (Wako, Tokyo, Japan), 2-(N-morpholino)ethane sulfonic acid (Dojindo Laboratories, Tokyo, Japan), and d -camphor-10-sulfonic acid (Wako) each at 20 μM]. The mixture was combined with four 3-mm zirconia beads (BioSpec Products, Bartlesville, OK, USA) and vigorously shaken for 5 min using a Shake Master NEO (Biomedical Science, Tokyo, Japan). Subsequently, deionized water (200 μl) and chloroform (500 μl) were added, and the suspension was vigorously shaken for 5 min and centrifuged at 4600g and 4°C for 30 min. The supernatant was filtered through a 5-kDa-cutoff filter column (Millipore, Burlington, MA, USA). The filtrate was concentrated via centrifugation (3 hours at 40°C) and dissolved in water containing reference compounds [200 μM each of 3-aminopyrrolidine (Sigma-Aldrich, St. Louis, MO, USA) and trimesic acid (Wako)] before CE-TOFMS analysis. As described previously (68 (link)), the CE-TOFMS experiments were performed using an Agilent CE-TOFMS system (Agilent Technologies).
5
Melanoma Metabolite Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells from 10 cm dishes were washed twice with 10 ml 5% mannitol in MilliQ water before metabolites were extracted from melanoma cell pellets using 1 ml methanol for 10 min and samples were deproteinized using 400 µl CHCl3 and 200 ml MilliQ water followed by centrifugation at 10,000 g for 3 min at 4°C. 400 µl of the aqueous layer was then filtered using a 5 kDa ultrafiltration tube and analysed by capillary electrophoresis mass spectrometry (CE‐MS) after addition of 25 µl 200 mM internal standards: L‐methionine sulfone (Wako 502–76641), 2‐(N‐morpholino) ethanesulfonic acid (Dojindo 349–01623), D‐Camphor‐10‐sulfonic acid (Wako 037–01032), 3‐aminopyrrolidine (Aldrich 404624) and trimesate (Wako 206–03641) as described (Kami et al., 2013 ) on an Agilent capillary electrophoresis system consisting of an Agilent G6220A LC/MSD TOF, an Agilent 1100 series isocratic HPLC pump, a G1603A Agilent CE‐MS adapter kit, and a G1607A Agilent CE‐ESI‐MS sprayer kit (Agilent Technologies, USA).
6
Decabromodiphenyl Ether Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Methanol, N,N-diethyl-2-phenylacetamide, D-camphor-10-sulfonic acid, and acetonitrile were purchased from Wako Pure Chemical Industries (Osaka, Japan). Ultrapure water was purified using a RFD280NC system (ADVAN-TEC, Dublin, CA, USA). BDE-209 and all other reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA), as described previously (Miyaso et al., 2012 (Miyaso et al., , 2014)) .
7
Endothelial Cell Culture and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following reagents were used: endothelial cell basal medium (EBM-2) and EGM-2 Single Quots (supplemental factors) (Lonza Japan Ltd., Tokyo, Japan); Dulbecco's modified Eagle's medium (DMEM), penicillin/streptomycin (100 ×), and trypsin-EDTA (Invitrogen/GIBCO, Grand Island, NY, USA); tissue-culture treated cell culture dishes/plates (Corning Inc., Tokyo, Japan); collagenase, trypsin inhibitor, mannitol, D-camphor-10-sulfonic acid (CSA), 1,3,5-benzenetricarboxylic acid (Trimesate), and gelatin (Wako Pure Chemical Industries Ltd., Osaka, Japan); 2-(N-morpholino) ethanesulfonic acid (MES) (Dojindo, Kumamoto, Japan); growth factor reduced (GFR) Engelbreth-Holm-Swarm (EHS) gel (BD Biosciences, Bedford, MA, USA); and acetaminophen (p-Acetamidophenol) (Nacalai Tesque Inc., Kyoto, Japan). All reagents used in the present study were of analytical grade.
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!