Starch was extracted and purified from 0.5g of malt flour using a modified version of the protocol described by Carciofi et al. (2011) . The residue after SDS and dithiothreitol (DTT) treatment was subjected to enzyme treatments, 2.5U of proteinase K (Sigma-Aldrich) and 0.1U of β-glucanase from Megazyme International Ltd, to remove protein and cell wall debris. Samples were digested overnight and filtered through a 100 μm pore size nylon filter. Crude starch slurry was centrifuged for 10min at 4000 g (Allegra X-22R centrifuge, Beckman Coulter, Germany). The precipitate was suspended in 200 μl of water, layered over 1ml of 80% (w/v) caesium chloride solution, and centrifuged at 13 000 g (Allegra X-22R centrifuge, Beckman Coulter) for 15min. The starch pellet was washed twice with water, followed by acetone washing and overnight air drying.
Allegra x 22r centrifuge
Manufactured by Beckman Coulter
Sourced in United States, Canada, Germany
The Allegra X-22R centrifuge is a high-performance benchtop centrifuge designed for a wide range of applications. It features a maximum speed of 22,000 rpm and a maximum RCF of 56,000 x g, providing efficient separation of samples. The Allegra X-22R is equipped with a microprocessor-controlled drive system and offers user-friendly programming options for precise control of run parameters.
Automatically generated - may contain errors
Lab products found in correlation
Desk centrifuge 5418, by Eppendorf (2 mentions)
0.2 μm nylon membrane, by Corning (2 mentions)
Centrifuge 5415r, by Eppendorf (2 mentions)
Proteinase k, by Merck Group (1 mentions)
β glucanase, by Megazyme (1 mentions)
15 ml centrifuge tubes, by BD (1 mentions)
Evos m7000 imaging system, by Thermo Fisher Scientific (1 mentions)
Eclipse ts100, by Nikon (1 mentions)
Cellasic onix2, by Merck Group (1 mentions)
Granulocyte macrophage colony stimulating factor gm csf, by Merck Group (1 mentions)
Ficoll histopaque 1077, by Merck Group (1 mentions)
Ampurex, by Beckman Coulter (1 mentions)
Nanodrop nd 1000, by Thermo Fisher Scientific (1 mentions)
Wizard genomic dna purification kit, by Promega (1 mentions)
Qubit fluorometer, by Thermo Fisher Scientific (1 mentions)
Cyan adp analyzer, by Beckman Coulter (1 mentions)
Fitc annexin 5 dead cell apoptosis kit, by Thermo Fisher Scientific (1 mentions)
Powerwave xs spectrophotometer, by Agilent Technologies (1 mentions)
Bio plex handheld magnetic washer, by Bio-Rad (1 mentions)
Oscillator, by Thermo Fisher Scientific (1 mentions)
28 protocols using allegra x 22r centrifuge
1
Starch Extraction and Purification from Malt Flour
2
Enveloped Virus Concentration Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Although several methods of concentration of samples were initially evaluated, the adsorption-precipitation protocol with aluminum chloride (AlCl3), previously described in Randazzo et al. (2019) (link), was selected because of better quality and quantity of RNA obtained. Briefly, 150 mL of water was transferred into a beaker of 200 mL and 75 μL of the enveloped rhabdovirus SVCV (105 TCID50/mL) (an enveloped RNA virus) was inoculated to each water sample as a concentration control. The SVCV virus was selected because it is an enveloped virus like the SARS-CoV-2, and it is frequently used in our research group. The pH of each sample was adjusted to 6.0 and 0.9 N AlCl3 solution was added to the sample at 1:100 ratio. The pH was again readjusted to 6.0 and samples were mixed at room temperature in an orbital shaker at 150 rpm during 15 min. Then, samples were centrifuged at 1700g for 20 min. in a Sorval ST Plus Series centrifuge (Thermo Scientific, USA) and the pellet was resuspended in 10 mL of 3% beef extract at pH 7.5 and transferred to 15 mL centrifuge tubes (Falcon). Samples were mixed again at room temperature in an orbital shaker at 200 rpm for 10 min. and centrifuged at 1900g for 30 min. in the Allegra™ X-22R Centrifuge (Beckman Coulter). Finally, the pellet was resuspended in 1 mL 1× PBS and samples were stored at −20 °C until RNA isolation.
3
Microfluidic Cell Chip Analysis Platform
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The microfluidic cell chip culture analysis platform CellASIC ONIX2 was purchased from Merck & Co., Inc. (Darmstadt, Germany); the EVOS M7000 imaging system was purchased from Invitrogen Life Technology Co., Ltd. (Carlsbad, CA, USA); the research-grade fluorescence inverted microscope ECLIPSE TS100 was purchased from Nikon Corporation (Nikon, Japan); the Allegra x-22R Centrifuge was purchased from Beckman coulter, Inc. (Brea, CA, USA); the MCO-18AIC CO2 Incubator was purchased from Panasonic Corporation (Tokyo, Japan); and the SpectraMax I3X Enzyme marker was purchased from Molecular Devices Instruments Ltd. (San Jose, CA, USA).
4
Freezing and Storage of Fenugreek Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Extracted FG samples were stored at 4 °C for eight days in total. At zero, one, two, four, and eight days, a small portion of each sample (400 mL) was collected and frozen at −28 °C until analyzed. For some analyses, FG was first precipitated by anhydrous ethanol with a ratio of 1:3 (FG extract:ethanol) followed by centrifugation (Allegra X-22R Centrifuge, Beckman Coulter, Mississauga, ON, Canada). Subsequently, the pellet was freeze-dried to obtain dried FG.
5
Isolation of Human Monocyte-Derived Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human monocyte-derived macrophages (HMDM) were isolated from the peripheral blood of healthy donors as previously described. Briefly, blood was layered at a ratio of 2:1 (blood/Ficoll medium) on Ficoll Histopaque-1077 (Sigma) in 15 ml centrifuge tubes and spun for 30 min at 2000 rpm in an Allegra X-22R centrifuge (Beckman Coulter). The layer containing the peripheral blood mononuclear cells was collected and then resuspended in 15 ml of PBS, and recentrifuged for 10 min at 1000 rpm. After two washes in PBS, cells were resuspended in DMEM containing 10% FBS, L-Glutamine and 100 units ml−1 penicillin and 100 mg ml−1 streptomycin on 12 mm diameter coverslips in 24-well plates. Non-adherent cells were removed after 4 h. The cells were subsequently cultured in cell culture medium containing 50 ng ml−1 granulocyte macrophage colony stimulating factor (GM-CSF) (Sigma Aldrich) in an atmosphere containing 5% CO2. Cultures were fed daily, and infection experiments were performed 10 days after the peripheral blood was collected. Infections were performed with MOI of 100:1:1 (bacteria/neutrophil/macrophage) ratio.
6
Piglet Growth and Muscle Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Body weights of piglets were recorded after an overnight fast. Before slaughter, plasma was collected into 10 mL tubes from the jugular vein for IL-15 determination. Serum was separated by centrifugation at 827 rcf (Beckman Coulter, Allegra X-22R Centrifuge, made in USA) for 15 min at 4 °C and then stored at −80 °C until analysis. The piglets were electrically stunned, exsanguinated and eviscerated. Immediately, samples (about 5 g) of the longissimus lumborum muscle dissected from the carcasses were placed in 10% neutral buffered formalin, or in liquid N2 and then stored at −80 °C, respectively, until further analyses.
7
Isolation and Preparation of Blood Components
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood was collected from healthy and consenting donors at the Centre for Blood Research with protocol approval from the University of British Columbia clinical ethics committee. Whole blood was collected in 3.8% sodium citrate coated tubes (BD VacutainerTM buffered citrate sodium (0.105 M; 9:1 blood/anticoagulant)). Serum was collected in silica spray coated serum tubes (BD VacutainerTM Plus Plastic Serum) and prepared by leaving tubes undisturbed to allow for clot formation for 30 min at room temperature, followed by centrifugation at 2000 g for 15 min in an Allegra X-22R centrifuge (Beckman Coulter, Canada). Platelet rich plasma (PRP) was collected by centrifuging citrated whole blood samples at 150 g for 12 min. Platelet poor plasma (PPP) was collected by centrifuging whole blood samples at 2000 g for 15 min. Red blood cell (RBC) suspensions were prepared by washing packed red cells with PBS for four times and resuspension in PBS to yield a 20% hematocrit cell suspension. We followed our published protocols to study the blood compatibility of the SMPNs20 (link),36 (link).
8
Bacterial DNA Extraction and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells from 20 mL of BPW cultures were collected by centrifugation at 1700 × g for 30 minutes at 4°C on Allegra X-22R centrifuge with SX4250 swing bucket rotor (Beckman Coulter, USA). Genomic DNA was extracted from the cell pellets with the Wizard Genomic DNA Purification Kit (Promega, Madison, WI). After resuspension, one half of the cells were processed with the manufacturer’s recommended protocol for Gram positive bacteria while the other half were processed with the protocol for Gram negative bacteria. The DNA concentration was measured by Qubit fluorometer (Invitrogen, USA) and the quality was determined by microspectrophotometry (NanoDrop ND-1000, NanoDrop technologies, Wilmington, DE). For each sampling occasion, equal volumes of Gram positive and Gram negative bacterial DNA extracts were mixed and cleaned by 1× SPRI beads (AMPureX, Beckman Coulter, CA, USA).
9
Neutrophil Plasma Membrane Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
First, 20 mL of whole blood was collected from male Sprague–Dawley rats by cutting femoral artery. Next, naïve neutrophils were collected from using a neutrophil collection kit (Haoyang TBD science, Tianjin, China) per the manufacturer׳s protocol. In brief, cells were suspended in 10 mL hypotonic lysis buffer. The buffer was consisted of potassium chloride (KCl), magnesium chloride (MgCl2), as well as one ethylene diamine tetraacetic acid (EDTA)-free mini protease inhibitor tablet (Selleck, Shanghai, China), and further broken using a homogenizer. The entire solution underwent 25 passes before spinning down at 3200×g (Allegra X-22R centrifuge, Beckman Coulter, USA), 4 °C, for 6 min. The supernatant was saved while the pellet was resuspended in lysing buffer and underwent another 20 passes and spun down again. The supernatants were pooled and centrifuged at 20,000×g, 4 °C, for 20 min, after which the pellet was thrown away and the supernatant was centrifuged again at 100,000×g, 4 °C, for 2 min. The pellet containing the plasma membrane was washed with 10 mmol/L pH7.5. Tris(hydroxymethyl)aminomethane-hydrogen chloride (Tris–HCl) and 1 mmol/L EDTA. The final pellet was collected and to be purified neutrophil plasma membrane for the following experiments.
10
Annexin V Assay for Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess phosphatidylserine translocation to the outer leaflet of the plasma membrane as a marker of apoptotic cell death, the FITC Annexin V/Dead Cell Apoptosis Kit (Invitrogen) was used following manufacturer’s instructions. Briefly, 1x106 cells were harvested, washed in ice cold PBS and pelleted by centrifugation at 500xg in an Allegra X-22R centrifuge (Beckman Coulter). Cells were resuspended in 1X annexin-binding buffer supplied by the kit, and sample was stained for 15 min at room temperature. At the end of the incubation, cells were diluted in additional 1X annexin-binding buffer and kept on ice. Staining was analyzed by flow cytometry using a CyAn ADP analyzer (Beckman Coulter) with fluorescence excitation and emission at 530nm and 575nm, respectively. Data were analyzed using FlowJo v. 10 software (FlowJo LLC, Ashland, OR).
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!