Vanillate
Manufactured by Merck Group
Vanillate is a laboratory equipment product manufactured by Merck Group. It functions as a device for the detection and measurement of vanillin, a common flavor compound. The core function of Vanillate is to provide accurate and reliable analysis of vanillin content in various samples.
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Lab products found in correlation
Guaiacol, by Merck Group (2 mentions)
Glutathione (gsh), by Merck Group (2 mentions)
Erythro gge, by Tokyo Chemical Industry (2 mentions)
L arginine, by Merck Group (1 mentions)
Mgso4, by Merck Group (1 mentions)
Ethyl acetate, by Thermo Fisher Scientific (1 mentions)
2 3 dichloro 5 6 dicyano p benzoquinone ddq, by Merck Group (1 mentions)
Lsm 800, by Zeiss (1 mentions)
Prolong diamond antifade mountant with dapi reagent, by Thermo Fisher Scientific (1 mentions)
Airyscan, by Zeiss (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 protocols using vanillate
1
Synthesis and Characterization of Novel Compounds
2
Antimicrobial Susceptibility Testing of Pseudomonas aeruginosa
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MICs were determined by broth microdilution following the guidelines from the Clinical and Laboratory Standards Institute and using P. aeruginosa ATCC27853 as a quality control. Susceptibility was assessed according to the interpretive criteria of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) or using a cut-off value of 16 mg/L for temocillin (no EUCAST breakpoint) by analogy with the current susceptibility breakpoint of its parent compound ticarcillin. For specific isolates, MICs were also measured in the presence of 20 mg/L Phe-Arg-β-naphthylamide dihydrochloride (PAβN, broad-spectrum efflux pump inhibitor10 (link); Sigma-Aldrich) and of 1 mM MgSO4 (to prevent the outer membrane permeabilization caused by PAβN33 (link)) or in the presence of substrates of P. aeruginosa porins, namely vanillate (VNL, OpdK substrate18 (link)), or L-arginine (L-Arg, OprD substrate19 (link)), both obtained from Sigma-Aldrich.
3
Synthesis and Purification of MPHPV and HPV
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Vanillate, guaiacol, GSH, and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) were purchased from Sigma-Aldrich. erythro-GGE was purchased from TCI America (Portland, OR).
A racemic mixture of MPHPV was synthesized by dissolving erythro-GGE into ethyl acetate (Fisher) and then slowly adding 1.25 molar eq of DDQ and stirring for 30 min. The reaction was washed three times with saturated NaHCO3 to remove DDQH2 formed during the reaction. The MPHPV was purified via flash chromatography using hexane/ethyl acetate (0.33:0.67, v/v), as described previously (15 (link)), and then crystallized from the eluent via solvent evaporation.
HPV was synthesized as described previously for synthesis of β-deoxy-α-veratrylglycerone, except using 4-O-benzyl-acetovanillone as starting material, rather than acetoveratrone (15 (link)). Synthesis of HPV required an additional debenzylation step that was unnecessary in the synthesis of β-deoxy-α-veratrylglycerone.
A racemic mixture of MPHPV was synthesized by dissolving erythro-GGE into ethyl acetate (Fisher) and then slowly adding 1.25 molar eq of DDQ and stirring for 30 min. The reaction was washed three times with saturated NaHCO3 to remove DDQH2 formed during the reaction. The MPHPV was purified via flash chromatography using hexane/ethyl acetate (0.33:0.67, v/v), as described previously (15 (link)), and then crystallized from the eluent via solvent evaporation.
HPV was synthesized as described previously for synthesis of β-deoxy-α-veratrylglycerone, except using 4-O-benzyl-acetovanillone as starting material, rather than acetoveratrone (15 (link)). Synthesis of HPV required an additional debenzylation step that was unnecessary in the synthesis of β-deoxy-α-veratrylglycerone.
4
FrankenSphingomonas Gene Expression Analysis
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FrankenSphingomonas 50 mL cultures in which gene expression was induced by addition of 250 μM Vanillate (4-hydroxy-3-methoxybezoic acid, Sigma-Aldrich) were growth until an optimum optical density was obtained (O.D. 600 nm = 0.6–0.7). Then, bacteria culture was centrifuged at 5,000 rpm for 5 min and the bacterial pellet resuspended in PBS. Cytospin prepation was performed and cells were fixed using 4% paraformaldehyde for 10 minat RT. After washing 3 times, samples were permeabilized with 0.15% Triton X-100 (Sigma-Aldrich) and washed again. Slides were blocked with 2% BSA (Sigma-Aldrich) in PBS-Tween 20 during 1 hat RT. The slides were then incubated overnight at 4°C with antibodies and protocol used for flow cytometry. After two washes with PBS the slides were mounted on ProLong Diamond Antifade Mountant with DAPI reagent (Thermo Scientific, Madrid, Spain). The sections were examined using a Zeiss LSM 800 with Airyscan (Carl Zeiss, Oberkochen, Germany).
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