The organic acid quantification was based on Rawi et al. (2021) (link). The fermentation sample from each sampling period was pipetted into a 2 mL microcentrifuge tube for centrifugation (Centrifuge-5804, Eppendorf) at 13000 rpm for 10 min to obtain a clear supernatant. The supernatant was filtered through a 0.22 μm syringe filter unit (Millipore) into an HPLC vial (Agilent Technologies, Cheshire, United Kingdom). Prominence Series Liquid Chromatography (Shimadzu Corp., Japan) using a reverse phase ion-exclusion C12 column (Rezex ROA, Phenomenex) was used for SCFA analysis. Analytes were detected using a UV detector at a wavelength of 210 nm. The isocratic mobile phase used was 0.25 mM sulphuric acid (H2SO4). A 15 μL sample was injected into the heated column (40°C) programmed to run in isocratic elution at a flow rate of 0.5 mL/min for 40 min. The peaks and response factors within the sample were calibrated and calculated using the LC Solutions software (Shimadzu). The standard solution contained of 12.5, 25, 50, 75, 100, 125, 150, 175, and 200 mM acetate, butyrate, propionate, and lactate.
0.22 μm syringe filter unit
Manufactured by Merck Group
Sourced in United Kingdom, United States
The 0.22 μm syringe filter unit is a laboratory equipment designed to remove particulates and microorganisms from fluid samples. It features a 0.22 μm pore size membrane that effectively retains bacteria and other contaminants, while allowing the desired fluid to pass through. This filter unit can be directly attached to a syringe for convenient sample filtration.
Automatically generated - may contain errors
Lab products found in correlation
Centrifuge 5804, by Eppendorf (2 mentions)
Hplc vial, by Agilent Technologies (2 mentions)
Lc solutions software, by Shimadzu (2 mentions)
Rezex roa, by Phenomenex (2 mentions)
Papain, by Merck Group (2 mentions)
Papain, by Worthington (2 mentions)
Uric acid, by Merck Group (1 mentions)
Huvecs, by ScienCell (1 mentions)
6 protocols using 0.22 μm syringe filter unit
1
HPLC Quantification of Organic Acids
2
Papain and 2W1S Peptide Treatment for Immune Modulation
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Papain and 2W1S Peptide Treatment
Mice were treated intranasally with 10 μg Papain (Worthington)
and 50 μg 2W1S peptide (peptide sequence: EAWGALANWAVDSA, GenScript)
diluted in sterile PBS on Day 0, Day 1 and Day 15, the mice were then euthanized
on Day 20 for analysis. The I-A(b) mouse 2W1S tetramer was kindly provided by
NIH Tetramer Core Facility. In some assays Papain was first diluted in sterile
PBS and filtered through 0.22 μm syringe filter unit (Millipore), the
sterile Papain was then intranasally administered to mice on Days 0, 2, 4, 6, 8,
10 and euthanized on Day 11 for analysis.
Mice were treated intranasally with 10 μg Papain (Worthington)
and 50 μg 2W1S peptide (peptide sequence: EAWGALANWAVDSA, GenScript)
diluted in sterile PBS on Day 0, Day 1 and Day 15, the mice were then euthanized
on Day 20 for analysis. The I-A(b) mouse 2W1S tetramer was kindly provided by
NIH Tetramer Core Facility. In some assays Papain was first diluted in sterile
PBS and filtered through 0.22 μm syringe filter unit (Millipore), the
sterile Papain was then intranasally administered to mice on Days 0, 2, 4, 6, 8,
10 and euthanized on Day 11 for analysis.
3
Uric Acid Effects on Endothelial Cells
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HUVECs obtained from ScienCell Research Laboratories (USA) were cultured in supplemented EBM‐2 basal media (EndoGRO™VEGF Supplement kit. Millipore). Uric acid (Sigma, Saint Louis, USA) was dissolved in warmed serum‐free EBM‐2 medium and filtered with 0.22 μm syringe filter unit (Millipore) as previously described 18 . UA crystals were not detected during the cell experiments by polarizing microscopy. Cells were pre‐treated with phloretin for 30 min. before addition of UA.
4
SCFA Analysis in Fermentation Samples
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The fermentation
sample from each sampling period was pipetted into a 2 mL microcentrifuge
tube for centrifugation (Centrifuge-5804, Eppendorf) at 13,000 rpm for 10 min to obtain a clear supernatant. The supernatant
was filtered through a 0.22 μm syringe filter unit (Millipore) into an HPLC vial (Agilent Technologies, Cheshire,
UK).
A Prominence Series liquid chromatography instrument (Shimadzu Corp., Japan) with a reverse-phase ion-exclusion
C12 column (Rezex ROA, Phenomenex) was used for the
analysis of SCFAs. The analytes were read with a UV detector at 210
nm wavelength. The isocratic mobile phase used was 0.25 mM sulphuric
acid (H2SO4). An amount of 15 μL of sample was injected into
the heated column (40 °C) programmed to run in isocratic elution
at a flow rate of 0.5 mL/min for 40 min. The peaks and response factor
within the sample were calibrated and calculated using LC Solutions
software (Shimadzu). The standard solution contained
acetate, butyrate, propionate, and also lactate at a series concentration
of 12.5, 25, 50, 75, 100, 125, 150, 175, and 200 mM.
sample from each sampling period was pipetted into a 2 mL microcentrifuge
tube for centrifugation (Centrifuge-5804, Eppendorf) at 13,000 rpm for 10 min to obtain a clear supernatant. The supernatant
was filtered through a 0.22 μm syringe filter unit (Millipore) into an HPLC vial (Agilent Technologies, Cheshire,
UK).
A Prominence Series liquid chromatography instrument (Shimadzu Corp., Japan) with a reverse-phase ion-exclusion
C12 column (Rezex ROA, Phenomenex) was used for the
analysis of SCFAs. The analytes were read with a UV detector at 210
nm wavelength. The isocratic mobile phase used was 0.25 mM sulphuric
acid (H2SO4). An amount of 15 μL of sample was injected into
the heated column (40 °C) programmed to run in isocratic elution
at a flow rate of 0.5 mL/min for 40 min. The peaks and response factor
within the sample were calibrated and calculated using LC Solutions
software (Shimadzu). The standard solution contained
acetate, butyrate, propionate, and also lactate at a series concentration
of 12.5, 25, 50, 75, 100, 125, 150, 175, and 200 mM.
5
Papain and 2W1S Peptide Treatment for Immune Modulation
6
Virus Inhibition by L. reuteri
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Live L. reuteri Protectis bacteria were co-incubated with CA6, CA16, EV71 or CB2 virus in 2 % DMEM at 37 °C and CO2 for an hour. The bacteria-virus mixtures were then spun down at 4,000 x g for 5 min at 4 °C. The culture supernatant was filtered with 0.22 μm syringe filter unit (Millipore, United States) before viral titer determination in RD cells (section 2.4).
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