The silkworm cocoons (J108 strain) were obtained from Queen Sirikit Sericulture Center (Khon Kaen, Thailand) in January 2020. Cocoons were aged 42 days at collection. The silkworm cocoon was cut into small square pieces of 1 × 1 cm. Twenty-four grams of cocoon materials were added to 700 mL of purified water and autoclaved (Tomy-SX-700, Tokyo, Japan) at 120 °C, 2.5 bar for 60 min. After filtration through Whatman No. 1, the filtrate was freeze-dried at −85 °C, 0.595 bar (Labconco, Kansas, MO, USA) to obtain the sericin extract [18 (link)]. The sericin extract was stored at 4 °C before use. The protein content was determined by Bradford assay and molecular weight profiles were determined by polyacrylamide gel electrophoresis (SDS-PAGE) [18 (link)].
Sx 700
Manufactured by TOMY
Sourced in Japan
The SX-700 is a versatile laboratory equipment designed for precise measurements and analysis. It features high-precision sensing capabilities and a robust construction for reliable performance in various laboratory settings.
Automatically generated - may contain errors
5 protocols using sx 700
1
Sericin Extraction from Silkworm Cocoons
2
Sericin and Purple Waxy Corn Extraction
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The sericin and purple waxy corn cob extractions were performed, as previously described [13 (link)]. The sericin from silkworm cocoons (J108 strain) was obtained from Queen Sirikit Sericulture Center (Khon Kaen, Thailand) in January 2020. Sericin was extracted by autoclave. The cocoons were aged 42 days at collection. The silkworm cocoon was cut into small square pieces of 1 × 1 cm. Twenty-four grams of the cocoon material was added to 700 mL of purified water and autoclaved (Tomy-SX-700, Tokyo, Japan) at 120 °C, 2.5 bar for 60 min. Then, the mixture was filtered through a Whatman No. 1 filter. The filtrate was then freeze-dried at −85 °C, 0.595 bar (Labconco, Kansas, MO, USA), to obtain the sericin extract.
The purple waxy corn cobs (Thai purple waxy corn) were obtained from the Plant Breeding Research Center for Sustainable Agriculture (Khon Kaen University, Thailand) and extracted by maceration in 50% ethanol (ratio of powder to solvent 1:25), with stirring at 25 °C for 24 h. Then, the filtrate was collected. The residue was then reextracted twice following the same procedure. The filtrates were pooled, evaporated, and then freeze-dried at −85 °C, 0.595 bar (Labconco), to obtain the crude extract (purple waxy corn cob extract, PWCC) [13 (link)]. Both sericin and PWCC were stored and protected from the light at −20 °C.
The purple waxy corn cobs (Thai purple waxy corn) were obtained from the Plant Breeding Research Center for Sustainable Agriculture (Khon Kaen University, Thailand) and extracted by maceration in 50% ethanol (ratio of powder to solvent 1:25), with stirring at 25 °C for 24 h. Then, the filtrate was collected. The residue was then reextracted twice following the same procedure. The filtrates were pooled, evaporated, and then freeze-dried at −85 °C, 0.595 bar (Labconco), to obtain the crude extract (purple waxy corn cob extract, PWCC) [13 (link)]. Both sericin and PWCC were stored and protected from the light at −20 °C.
3
Synthesis and Characterization Protocol
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A
hot-air oven (Binder,
FED 53, Germany), an orbital shaker (GFL, 3020, Germany), a rotary
evaporator (BUCHI, RE-111 Rotavapor, Switzerland), a freeze dryer
and vacuum concentrator (LaboGene, Scanvac, Denmark), and a hot plate
stirrer (Ingenieurbüro CAT, M. Zipperer GmbH, M 6, Germany)
were used for material synthesis. For all aseptic experiments, an
autoclave (TOMY, SX-700, Japan) and a laminar flow cabinet (BossTech,
HVT, Thailand) were used. In all batch experiments, an incubator shaker
(New Brunswick, Innova 42) was used to investigate the effects of
dose, contact time, pH, and concentration, whereas a jar test (Wizard,
Plus 6, Thailand) was used for adsorption kinetics. Finally, an incubator
(Termaks, KBP 6087, Norway) was used to incubate sample tests in agar
diffusion tests, antibacterial batch experiments, adsorption isotherm,
kinetics, and desorption experiments.
hot-air oven (Binder,
FED 53, Germany), an orbital shaker (GFL, 3020, Germany), a rotary
evaporator (BUCHI, RE-111 Rotavapor, Switzerland), a freeze dryer
and vacuum concentrator (LaboGene, Scanvac, Denmark), and a hot plate
stirrer (Ingenieurbüro CAT, M. Zipperer GmbH, M 6, Germany)
were used for material synthesis. For all aseptic experiments, an
autoclave (TOMY, SX-700, Japan) and a laminar flow cabinet (BossTech,
HVT, Thailand) were used. In all batch experiments, an incubator shaker
(New Brunswick, Innova 42) was used to investigate the effects of
dose, contact time, pH, and concentration, whereas a jar test (Wizard,
Plus 6, Thailand) was used for adsorption kinetics. Finally, an incubator
(Termaks, KBP 6087, Norway) was used to incubate sample tests in agar
diffusion tests, antibacterial batch experiments, adsorption isotherm,
kinetics, and desorption experiments.
4
Immunohistochemical Analysis of Formalin-Fixed Paraffin-Embedded Tissues
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All formalin-fixed and paraffin-embedded tissues were obtained from the archives of the Department of Dermatology at Kaohsiung Chang Gung Memorial Hospital, Taiwan. First, 5 µm paraffin sections were deparaffinized by xylene and rehydrated with 100%, 95%, 75%, 50% ethanol, and water. Sections were heated in an autoclave (SX-700, TOMY, Tokyo, Japan) for 5 min in 10 mM citric acid buffer with 0.05% Tween-20. Sections were allowed to cool down. After rinsing with water, sections were blocked with 3% BSA in PBS for 1 h at room temperature. Sections were incubated with primary antibodies overnight at 4 °C in a humid chamber. After being washed with 0.05% Tween-20 PBS, sections were incubated with secondary antibodies for 1 h at room temperature. Nuclei were counterstained with DAPI (D9542, Sigma-Aldrich, St. Louis, MO, USA) and mounted with mounting media. Stained sections were examined using the BX53 microscope equipped with a DP80 camera (Olympus, Tokyo, Japan).
5
Microbial Concentration Preparation
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S. aureus (DMST
562) and E. coli (DMST 4212) (DMST
stands for Department of Medical Sciences Culture Collection, Thailand)
were used in this study. The water samples were prepared by diluting S. aureus and E. coli of 108 CFU/mL in sterile deionized water (autoclave:
TOMY, SX-700, Japan) to obtain initial S. aureus and E. coli concentrations from 104 to 107 CFU/mL.Note: CFU stands for
colony-forming unit, which is widely used to estimate the number of
visible microorganisms in test samples.91 (link)
562) and E. coli (DMST 4212) (DMST
stands for Department of Medical Sciences Culture Collection, Thailand)
were used in this study. The water samples were prepared by diluting S. aureus and E. coli of 108 CFU/mL in sterile deionized water (autoclave:
TOMY, SX-700, Japan) to obtain initial S. aureus and E. coli concentrations from 104 to 107 CFU/mL.
colony-forming unit, which is widely used to estimate the number of
visible microorganisms in test samples.91 (link)
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