Defatted or low-fat soybean flour samples (3.00 ± 0.01 g) and distilled water (1:10 w/v) were added into a 50 ml centrifuge tube and mixed in a shaking incubator (Incu-Shaker Mini, Benchmark Scientific; Sayreville, NJ, United States) at 250 rpm, at two different temperatures (22 and 60°C) for two different durations (30 and 60 min). In the first trial, the 30 min duration was tested at 22°C and 60°C, respectively. Because the increased extraction temperature did not increase SPC protein content substantially, not to mention of its higher energy consumption, for the 60 min extraction, only 22°C was examined in the next trial. The slurry was then centrifuged at 3,810 × g for 30 min at room temperature (Sorvall™ ST 8, Thermo Scientific™; Waltham, MA, United States). After the solid–liquid separation, the precipitate (aka wet protein concentrate) was transferred into a pre-weighed drying pan for drying in a conventional lab oven (Heratherm™, Thermo Scientific™, Waltham, MA United States) at 60°C for 2.5 h. Dried SPC (contained 10–15% moisture) was ground into powder (to pass 100 μm openings) using a coffee grinder (Hamilton Beach Inc.; Southern Pines, NC, United States). Ground SPC samples and raw materials were kept at-20°C until further analysis.
Sorvall st8
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The Sorvall ST8 is a general-purpose, compact, and high-performance centrifuge designed for a wide range of laboratory applications. It offers fast acceleration and deceleration, and can accommodate various rotor types to handle different sample sizes and volumes.
Automatically generated - may contain errors
Lab products found in correlation
Incu shaker mini, by Benchmark Scientific (1 mentions)
Mrs broth medium, by Hopebio (1 mentions)
Blpr2 pressure transducer, by World Precision Instruments (1 mentions)
Polyethylene tubing, by BD (1 mentions)
Raw264, by American Type Culture Collection (1 mentions)
Freezone 12, by Labconco (1 mentions)
Ni nta resin, by Smart-Lifesciences (1 mentions)
Corneal epithelial cell growth kit, by American Type Culture Collection (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Pcs 420 032, by American Type Culture Collection (1 mentions)
Penicillin streptomycin amphotericin b solution, by American Type Culture Collection (1 mentions)
Prostate epithelial basal medium, by American Type Culture Collection (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Corning (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
T75 tissue culture flask, by Greiner (1 mentions)
Ficoll paque plus, by Merck Group (1 mentions)
Human msc phenotyping kit, by Miltenyi Biotec (1 mentions)
Nexera lc 30a system, by Shimadzu (1 mentions)
Tsb medium, by Hopebio (1 mentions)
11 protocols using sorvall st8
1
Soy Protein Concentrate Extraction
2
Yogurt Centrifugation Protocol for Syneresis and WHC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
15 g of yogurt samples were transferred to a 50 ml centrifuge tube to be centrifuged (Sorvall ST8; Thermo Scientific, Waltham, MA) for 10 min at 5000 xg and 4 °C. Three measurements were performed. The weight of supernatants and precipitates were recorded, and syneresis and WHC were calculated as follows:
3
Preparation of Lactobacillus plantarum M1 Cell-Free Supernatant
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The activated Lpb. plantarum M1 (OD600 approximately 1.6) was inoculated into MRS broth medium (Qingdao Hope Bio-technology Co., Ltd., Qingdao, China) at 2% (v/v) inoculum for 16 h at 37 °C and then centrifuged at 6200× g for 10 min (Sorvall ST8, ThermoFisher Scientific Co., Ltd., Shanghai, China) at 4 °C to collect the bacteria. The collected bacteria were added to 1/10 of the original volume of fresh MRS broth (At this point, the inoculum density is approximately 1010 CFU/mL) at 37 °C for 16 h, and then centrifuged at 6200× g for 10 min at 4 °C. The supernatant was filtered through a 0.22 μm sterile aqueous filter membrane to remove other debris to obtain a CFS.
4
Rodent Plasma Collection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty-four hr after the exposure, rats were anesthetized using isoflurane (5% induction and 3% maintenance). While anesthetized, animals were weighed, and placed in a nose cone on a surgical board in a supine position where the anesthetic was maintained. Once the animal was secured to the board, the carotid artery was isolated and cannulated with polyethylene tubing with an inner diameter of 0.58 mm and outer diameter of 0.965 mm (BD Intramedic, Franklin Lakes, NJ). MAP was obtained using a BLPR2 pressure transducer as well as a blood pressure monitor (World Precision Instruments, Sarasota, FL). After a stable MAP reading was recorded, approximately 6 mL of blood was collected directly from the cannula into BD Vacutainer® Plus whole blood tubes lined with 100% Dipotassium EDTA Dihydrate. Whole blood was centrifuged (Sorvall™ ST 8, Thermofisher Scientific, Waltham, MA) at 1100 RCF for 10 min (Thermofisher Scientific, Waltham, MA). The plasma was then removed using a pipette, aliquoted for future study, and snap frozen in liquid nitrogen.
5
Murine Monocyte and Macrophage Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Murine monocytes cell line (RAW 264.7) was obtained from American Type Culture Collection (Manassas, VA, USA). Cultured the cells in Dulbecco's modified Eagle's medium with 10% (v/v) FBS, 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C in a humidified atmosphere with 5% CO2 at 37 °C. Peritoneal macrophages were isolated from C57BL/6 J mice according to previously reported with some modifications19 (link). Briefly, harvested the cells by washing the peritoneal cavity with ice-cold PBS at 5 mL per mouse followed centrifuged at 1000 rpm (Sorvall ST 8, Thermo Fisher Scientific, Suzhou, China) for 10 min. Then, the cells were cultured in RPMI-1640 medium supplemented with 10% (v/v) fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin. These macrophages were seeded on 6-well plates and maintained in a water-saturated atmosphere 5% CO2 at 37 °C. Non adherent cells were removed at 4 h after seeding.
6
Defatting Cocoa Beans for Polyphenol Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A control sample was prepared by freeze-drying the beans that were dehydrated directly and applying them to a freeze-dryer apparatus (FreeZone 12, Labconco Corporation, Kansas City, MO, USA). After freeze-drying, the beans were milled in an ultracentrifuge knife-miller (ZM200, Retsch, Dusseldorf, Germany) until obtaining a particle size of 500 μm. Since in cocoa beans, the total fat content exceeds 50% on a dry basis [50 ], the fat fraction interferes in the extraction process, causing a low recovery of polyphenolic compounds. For this reason, a method was applied for defatting the sample before extraction. The defatting process was carried out by weighing 2 g of sample in centrifuge tubes and mixing them with 20 mL of hexane. The mixture was shaken for 30 min at 200 rpm in a horizontal shaker and subsequently centrifuged for 20 min at 8000 rpm in a centrifuge (Sorvall ST8, Thermo Scientific, Waltham, MA, USA). The sample was then recovered, and the supernatant was discarded. Finally, the samples were placed under a vacuum for 15 h to eliminate the residual hexane.
7
Recombinant Protein Expression and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tree TLP genes were expressed in the E. coli BL21 (DE3) strain. The cells grown at 37 °C with shaking at 180 rpm in 250 mL of LB broth, containing 50 μg/mL Ampicillin until an OD600 reached 0.8. The expression of the recombinant protein was induced by the addition of isopropyl β-D-1-thiogalactopyranoside (IPTG) at the final concentration of 0.1 mmol/L, followed by cultivation for 24 h at 15 °C with 160 rpm (High speed tabletop centrifuge, Sorvall ST8, Thermo Fisher Scientific Corporation, MA, USA). The culture was harvested by centrifugation (10,000×g for 15 min at 4 °C), and the cell pellets were re-suspended in 30 mL of buffer A (50 mmol/L Tris–HCl pH 8 and 100 mmol/L NaCl) and were incubated for 30 min on ice. The cells were disrupted by sonication, and the insoluble fraction was collected by centrifugation (14,400 rpm for 60 min at 4 °C).
The three protein purification was performed in a single gravity flow chromatography step using a column packed with 1 mL of Ni-NTA resin (smart-lifesciences, China) and was equilibrated with buffer A. The purified protein was eluted using a five-step gradient of imidazole (10, 20, 80 and 300 mmol/L) in buffer A, each step containing 10 mL of the respective buffer. The purity of the protein samples was estimated by SDS-PAGE and stained with Coomassie brilliant blue G-250.
The three protein purification was performed in a single gravity flow chromatography step using a column packed with 1 mL of Ni-NTA resin (smart-lifesciences, China) and was equilibrated with buffer A. The purified protein was eluted using a five-step gradient of imidazole (10, 20, 80 and 300 mmol/L) in buffer A, each step containing 10 mL of the respective buffer. The purity of the protein samples was estimated by SDS-PAGE and stained with Coomassie brilliant blue G-250.
8
Enzymatic Hydrolysis of Biomass
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
According to the literature [19 (link)], 20 FPU/g of cellulase (CAS: 9012-54-8) was added to assay the saccharification efficiency of the biomass before and after the lignin degradation treatment. After the saccharification reaction, samples were centrifugated with high-speed centrifuge (Sorvall ST8, ThermoFisher) at 5000 rpm for 3 min, and the reducing sugar content was determined simultaneously by HPLC and DNS method. The saccharification efficiency was calculated as follows: saccharification efficiency (or the yield of enzymatic hydrolysis) % = Mass of reducing sugar produced *0.9/mass of cellulose and hemicellulose in the sample.
9
Primary Bladder Epithelial Cell Expansion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary bladder epithelial cells (PCS-420-032, ATCC, Manassas, VA, USA) were purchased and expanded using the provided protocol. Briefly, cells were transferred to a T75 flask at 4000 cells per cm2 with Prostate Epithelial Basal Medium (ATCC, Manassas, VA, USA) supplemented with Corneal Epithelial Cell Growth Kit (ATCC, Manassas, VA, USA) and Penicillin–Streptomycin–Amphotericin B Solution (ATCC, Manassas, VA, USA) at 5 mL of complete growth media per 25 cm2. Every 48 h, spent medium was removed and fresh medium was added until approximately 80% confluence was reached. Upon reaching approximately 80% confluence, spent medium was removed and cell layer was detached using 0.05% Trypsin–EDTA (Gibco, Thermo Fisher Scientific, Waltham, MA, USA). Trypsin–EDTA was neutralized with equal volume fetal bovine serum (Corning, Corning, NY, USA). Cells were collected and centrifuged (Sorvall ST8, Thermo Fisher Scientific, Waltham, MA, USA) for 5 min at 150 g. Cells were counted and either frozen in working volumes with 10% dimethyl sulfoxide (Thermo Fisher Scientific, Waltham, MA, USA) or used for wound-healing assay at passage 4.
10
Isolation and Characterization of MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow in the transportation medium was overlaid on approximately 20 ml of sterile Ficoll-Paque Plus (Merck KGaA, Darmstadt, Germany) taken in a conical centrifuge tube (Sarstedt AG & Co, Nümbrecht, Germany) and centrifuged in a swing bucket centrifuge (Sorvall ST8, Thermo Fisher Scientific, Massachusetts, USA) at 2000 rpm for 5 minutes at 4°C. The resultant layer containing mononuclear cells were siphoned out and seeded onto a T75 tissue culture flask (Greiner Bio-One GmbH, Frickenhausen, Germany) at the rate of 5000 cells/cm [2 (link)]. Approximately 20 ml of the MSC culture medium (CTS™ StemPro™ MSC SFM, Thermo Fisher, Massachusetts, USA) was added, and the flask was incubated at 37 C, with 5% CO2 and 95% RH in a CO2 incubator. Adherent cells were cultured, and the process was continued with change of the medium every second day till cells became confluent. The cells were trypsinised, harvested, and used for chondrocyte differentiation. A portion of the cells was used for cell characterization using MSC specific antibodies in a flow cytometer (Human MSC Phenotyping Kit from Miltenyi Biotec (Auburn, CA, USA)). Positive expression of CD73, CD90, and CD105 and negative for CD45 were confirmed.
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!