A carnosic acid and carnosol standard as well as 2,2-Diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Other solvents were obtained from J.T. Baker (Radnor, PA, USA). All components for the preparation of eutectic solvents are commercially available from suppliers such as Sigma-Aldrich (St. Louis, MO, USA), Acros Organics (Waltham, MA, USA), and Gram mol (Zagreb, Croatia). Macroporous resins (XAD7, XAD4, XAD16) were purchased from Sigma-Aldrich (St. Louis, MO, USA) and HP21 and HP20 from Mitsubishi Chemical Holdings (Chiyoda City, Tokio, Japan). The characteristics of the macroporous resins, according to the manufacturer and other authors [30 (link),31 (link)], are shown in Table 1 .
Xad16
Manufactured by Merck Group
Sourced in United States, Germany
XAD16 is a macroporous non-ionic polymeric adsorbent resin. It has a high surface area and can be used for the adsorption and separation of a wide range of organic compounds from aqueous solutions.
Automatically generated - may contain errors
Lab products found in correlation
Amberlite xad 16 resin, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
Sodium hydroxide (naoh), by Avantor (1 mentions)
1 1 diphenyl 2 picrylhy drazyl dpph 2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Sinapic acid, by Merck Group (1 mentions)
6 protocols using xad16
1
Characterization of Macroporous Resins
2
Optimization of Microbial Metabolite Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Batch cultures were carried out in 250-mL Erlenmeyer flasks containing 50 mL medium with varying initial pH (5.5, 6.5, 7.0, 7.5 and 8.5). Ten percent (v/v) of the seed culture was used to inoculate the flasks. The culture was incubated on a rotary shaker at 28 °C and 150 rpm. After 72 h incubation, the cultures were centrifuged (12,000×g, 20 min, 4 °C) to separate the bacterial cells and the cell filtrate was stored at 4 °C until used. The cultures were vigorously extracted with equal volume of ethyl acetate three times, mixed, centrifuged, and the top organic layer was dried for analysis.
Liquid cultures were grown at 150 rpm on a shaker in 250 mL Erlenmeyer flasks containing 50 mL medium and 2% (v/v) of XAD-16 (Sigma-Aldrich). These cultures were inoculated with 10% (v/v) of a 24 h preculture in the same medium without XAD-16. Cultures were harvested after 72 h, and XAD beads were separated from the cells and supernatant by sieving. XAD beads were extracted with MeOH (25 mL), and the MeOH extract was concentrated to dryness on a rotary evaporator. The residue was re-dissolved in MeOH (1.5 mL) for HPLC/MS analysis.
Liquid cultures were grown at 150 rpm on a shaker in 250 mL Erlenmeyer flasks containing 50 mL medium and 2% (v/v) of XAD-16 (Sigma-Aldrich). These cultures were inoculated with 10% (v/v) of a 24 h preculture in the same medium without XAD-16. Cultures were harvested after 72 h, and XAD beads were separated from the cells and supernatant by sieving. XAD beads were extracted with MeOH (25 mL), and the MeOH extract was concentrated to dryness on a rotary evaporator. The residue was re-dissolved in MeOH (1.5 mL) for HPLC/MS analysis.
3
Isolation and Fractionation of Anti-Inflammatory Metabolites from Lactobacillus rhamnosus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To test whether the strain that exhibited the most anti-inflammatory activity mediated this action through secreted molecules, we tested the activity of the bacterial extract using the same experimental design. Three liters of the L. rhamnosus strain was grown in MRS media for 48 h without shaking; then, the produced metabolites were collected in a mixture of synthetic resin, including HP20, XAD7, and XAD16 (Sigma-Aldrich, St. Louis, MO, USA), with 60 g of resin per liter of the culture, and shaken for 2 h. Resin was collected via filtration, washed twice with water, and then eluted with methanol. The methanol extract was then concentrated to dryness, and the residue was redissolved in water/acetonitrile and fractionated using a Biotage automated chromatography system on reversed phase C18 silica using water/acetonitrile as a mobile phase (20:100%); this generated 10 fractions. In parallel, we created two other preparations via precipitation of the cell-free supernatant using ammonium sulfate to enrich the proteins. All 12 fractions were then tested for immunomodulatory activity.
4
Screening of Secondary Metabolites in M. gryphiswaldense
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the screening of secondary metabolites, M. gryphiswaldense and Δtrans-at-pks strains were cultivated at 28°C in FSM (33 (link)) with an initial OD at 565 nm (OD565) of 0.01 under aerobic, microoxic, and anaerobic conditions in 500-ml baffled Erlenmeyer flasks, in Duran Laboratory flasks with rubber stoppers containing 50 ml of medium, and in 250-ml Duran Laboratory flasks containing 240 ml of degassed medium with rubber stoppers, respectively. One milliliter (vol/vol) of sterile Amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) was added to the culture grown under aerobic and microoxic conditions and 5 ml (vol/vol) of XAD-16 into the culture grown under anaerobic conditions. The culture under aerobic condition was agitated at 150 rpm. The cells and the resin were harvested together by centrifugation after 60 h of incubation before extraction.
To access the activation of the cluster, wild-type, PmamDC45-trans-at-pks, and Δtrans-at-pks strains were cultivated under aerobic conditions at 28°C in 100 ml of FSM in a 1-liter baffled Erlenmeyer flask with a starting OD565 of 0.01 at 150 rpm. The culture was supplemented with 2 ml (vol/vol) sterile Amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany). After 60 h of incubation, the cells and resin were harvested together by centrifugation.
To access the activation of the cluster, wild-type, PmamDC45-trans-at-pks, and Δtrans-at-pks strains were cultivated under aerobic conditions at 28°C in 100 ml of FSM in a 1-liter baffled Erlenmeyer flask with a starting OD565 of 0.01 at 150 rpm. The culture was supplemented with 2 ml (vol/vol) sterile Amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany). After 60 h of incubation, the cells and resin were harvested together by centrifugation.
5
Rapeseed Meal Polyphenol Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The rapeseed meal was provided by Olead (Pessac, Bordeaux, France). Sinapine standard was purchased from ChemScience (JJ08852, Monmouth Junction, NJ, USA). Sinapic acid and formic acid (FA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium chloride (NaCl) and sodium hydroxide (NaOH) pellets were purchased from VWR (Radnor, PA, USA). Chlorohydric acid (HCl) 37% was from CarloErba (Val-de-Reuil, France). Acetonitrile (ACN) solution was provided by Biosolve BV (Valkensward, Netherlands). Absolute ethanol (EtOH) was purchased from DASIT (Paris, France). All solvents and chemical reagents used in this study were analytical and high-performance liquid chromatography (HPLC)-grade. 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium persulfate, and ascorbic acid (vitamin C) were provided by Sigma-Aldrich (St. Louis, MO, USA).
Five macroporous resins (XAD7, XAD4, XAD16, XAD1180, and HP20) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The characteristics of these macroporous resins are shown inTable 7 .
Five macroporous resins (XAD7, XAD4, XAD16, XAD1180, and HP20) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The characteristics of these macroporous resins are shown in
6
Screening and Activation of Trans-AT Polyketide Synthase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the screening of secondary metabolites, M. gryphiswaldense and ∆trans-at-pks strains were cultivated at 28°C in FSM medium (33) (link) with an initial OD565 of 0.01 under aerobic, microoxic and anaerobic conditions in 500 ml baffled Erlenmeyer flasks, and Duran Laboratory flasks with rubber-stopper containing 50 ml medium, and in 250ml Duran Laboratory flasks containing 240 ml degassed medium with rubber-stoppers, respectively. 1 mL (v/v) sterile amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) was added to the culture grown under aerobic and micro-oxic conditions and 5 ml (v/v) XAD-16 into the culture grown under anaerobic conditions. The culture under aerobic condition was agitated at 150 rpm. The cells and the resin were harvested together by centrifugation after 60 hours of incubation before extraction.
To access the activation of the cluster, wild type, PmamDC45-trans-at-pks, and ∆trans-at-pks strains were cultivated under aerobic condition at 28°C in 100 ml FSM medium in 1L baffled Erlenmeyer flask with starting OD565 of 0.01 at 150 rpm. The culture was supplemented with 2 ml (v/v) sterile amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany). After 60 hours of incubation, the cells and resin were harvested together by centrifugation.
To access the activation of the cluster, wild type, PmamDC45-trans-at-pks, and ∆trans-at-pks strains were cultivated under aerobic condition at 28°C in 100 ml FSM medium in 1L baffled Erlenmeyer flask with starting OD565 of 0.01 at 150 rpm. The culture was supplemented with 2 ml (v/v) sterile amberlite resin XAD-16 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany). After 60 hours of incubation, the cells and resin were harvested together by centrifugation.
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!