Seaprep agarose

Manufactured by Lonza

Sourced in United States, Switzerland

SeaPrep Agarose is a laboratory product designed for use in various applications. It functions as an agarose gel material, providing a medium for the separation and analysis of biomolecules such as DNA, RNA, and proteins. The core function of SeaPrep Agarose is to facilitate the electrophoretic separation and visualization of these biological samples.

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Lab products found in correlation

Bodipy 500 510 c4 c9, by Thermo Fisher Scientific (1 mentions) Rhodamine b, by Merck Group (1 mentions) Deuterium oxide, by Merck Group (1 mentions) Formalin, by Merck Group (1 mentions) Crystal violet solution, by Merck Group (1 mentions) Spectramax minimax 300 imaging cytometer, by Molecular Devices (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions) D300e digital dispenser, by Tecan (1 mentions) Zellshield, by Minerva Biolabs (1 mentions) Hepes, by Merck Group (1 mentions)

5 protocols using seaprep agarose

Composite Agarose Hydrogel for Nanoparticle Delivery

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Ultralow gelling temperature SeaPrep Agarose (Lonza) was dissolved in aCSF to a concentration of 3% (w/v) by microwaving on high power in 4-s bursts and vortexing until clear. The solution was allowed to cool to room temperature before further handling. Nanoparticles were dispersed in aCSF at 20% (w/w) by 5 min of bath sonication, and BDNF was then added. Room temperature agarose was added to the nanoparticle dispersion at a 1:1 volume ratio, and a dual asymmetric centrifugal mixer was used to create the composite agarose. One hundred microliters of composite agarose was pipetted into a 2-ml centrifuge tube and allowed to gel at 4°C for 1 hour.

Pakulska M.M., Elliott Donaghue I., Obermeyer J.M., Tuladhar A., McLaughlin C.K., Shendruk T.N, & Shoichet M.S. (2016). Encapsulation-free controlled release: Electrostatic adsorption eliminates the need for protein encapsulation in PLGA nanoparticles. Science Advances, 2(5), e1600519.

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Soy Protein Isolate Characterization

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CSPI (Fujipro-F), canola oil, medium-chain triglyceride (MCT, product name MCT64) oil, and rapeseed oil were supplied from Fuji Oil (Osaka, Japan). Fujipro-F is a standard commercial SPI that is untreated with any enzymatic or chemical modifications. Fujipro-F was heat-treated to 140 °C for 10 s for sterilization and powdered by spray-drying (Kawaguchi, Kita, Shinyashiki, Yagihara, & Fukuzaki, 2018 (link)). Deuterium oxide and Rhodamine B were purchased from Sigma-Aldrich (St. Louis, MO, US); BODIPY 500/510 C4 C9 was purchased from Thermo Fisher Scientific (Waltham, MA, US); Seaprep Agarose was purchased from Lonza (Basel, Switzerland); dialysis tubes (MWCO 3500 g/mol) were purchased from Nihon Medical Science (Osaka, Japan); and all other reagents were purchased from Kishida Chem (Osaka, Japan).

Kano H, & Shiraki K. (2023). Heat treatment in the presence of arginine increases the emulsifying properties of soy proteins. Food Chemistry: X, 17, 100567.

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Vero E6 Cell-based Plaque Assay

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Vero E6 cells were seeded in 24-well plates at a concentration of 7.5 × 104 cells/well. The following day, serial dilutions were performed in serum-free MEM media. After 1 h absorption at 37 °C, 2× overlay media was added to the inoculum to give a final concentration of 2% (v/v) FBS/MEM media and 0.4% (w/v) SeaPrep Agarose (Lonza) to achieve a semi-solid overlay. Plaque assays were incubated at 37 °C for 3 days. Samples were fixed using 4% Formalin (Sigma–Aldrich) and plaques were visualised using crystal Violet solution (Sigma–Aldrich).

Meyer B., Chiaravalli J., Gellenoncourt S., Brownridge P., Bryne D.P., Daly L.A., Grauslys A., Walter M., Agou F., Chakrabarti L.A., Craik C.S., Eyers C.E., Eyers P.A., Gambin Y., Jones A.R., Sierecki E., Verdin E., Vignuzzi M, & Emmott E. (2021). Characterising proteolysis during SARS-CoV-2 infection identifies viral cleavage sites and cellular targets with therapeutic potential. Nature Communications, 12, 5553.

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Measuring Anchorage-Dependent and -Independent Cell Growth

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To measure anchorage-dependent growth, cells were plated at low density in 96 well plates and allowed to attach overnight. Inhibitors were then added with a D300e Digital Dispenser (Tecan). After 72 hours, cells were labeled with calcein AM (Thermo Fisher) according to the manufacturer’s protocol, and counted on a SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices).
To measure anchorage-independent growth, soft agar colony formation assays were performed. In a 96-well plate, wells were coated with 50 μl 2% SeaPrep Agarose (Lonza) mixed with cell culture medium. Next, 100 μl of 5000 cells mixed with 1% SeaPrep Agarose were added to each well and allowed to solidify. Fifty μl of medium were added to each well, and drugs were dispensed with a D300e Digital Dispenser (Tecan). After 72 hours, the alamarBlue cell viability reagent (Thermo Fisher) was added and incubated for 3 hours. Fluorescence was quantified on a SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices).

Blake D.R., Vaseva A.V., Hodge R.G., Kline M.P., Gilbert T.S., Tyagi V., Huang D., Whiten G.C., Larson J.E., Wang X., Pearce K.H., Herring L.E., Graves L.M., Frye S.V., Emanuele M.J., Cox A.D, & Der C.J. (2019). Application of a MYC degradation screen identifies sensitivity to CDK9 inhibitors in KRAS-mutant pancreatic cancer. Science signaling, 12(590), eaav7259.

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Perfusion Culture of Cells in Agarose

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The pellet of cells after electroporation was mixed with 2% (w/w) SeaPrep agarose (Lonza, Switzerland) in DMEM at 37 °C, in v-v ratio 1:1. A HPLC capillary of 0.75 mm inner diameter was filled with the suspension and incubated on ice for 10 min. Upon gelling, a thread was pushed from the capillary into a 5 mm NMR tube filled with degassed bioreactor media: DMEM without NaHCO₃ (Sigma-Aldrich, USA) + 10% D₂O (Eurisotop, France) + 70 mM HEPES (Sigma-Aldrich, USA) + 1x ZellShield (Minerva Biolabs, Germany). Medium flow through the sample was set to 50 μl/min, using an HPLC pump-driven system that draws the medium from a reservoir heated in a water bath through a vacuum-degassing chamber and brings the fresh medium to the bottom of the NMR tube through a central glass capillary^{61 (link)}.

Víšková P., Ištvánková E., Ryneš J., Džatko Š., Loja T., Živković M.L., Rigo R., El-Khoury R., Serrano-Chacón I., Damha M.J., González C., Mergny J.L., Foldynová-Trantírková S, & Trantírek L. (2024). In-cell NMR suggests that DNA i-motif levels are strongly depleted in living human cells. Nature Communications, 15, 1992.

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