Ultracentrifuge xl 100 k

Manufactured by Beckman Coulter

Sourced in United States

The Ultracentrifuge XL-100 K is a high-speed centrifuge designed for the separation and analysis of macromolecules, such as proteins, nucleic acids, and subcellular organelles. It can achieve speeds up to 100,000 revolutions per minute (rpm) and can generate centrifugal forces up to 802,000 times the force of gravity. The Ultracentrifuge XL-100 K is a versatile laboratory instrument used in various fields, including biochemistry, molecular biology, and cell biology.

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Lab products found in correlation

Calcium phosphate transfection kit, by Takara Bio (2 mentions) Tecnai 12 electron microscope, by Thermo Fisher Scientific (1 mentions) 175 cm2 cell culture flasks, by Corning (1 mentions) Nanosight ns300, by Malvern Panalytical (1 mentions)

Spelling variants of this product

Ultracentrifuge (302 citations) XL-100 (2 citations) Beckman Optima XL-100 K Ultracentrifuge (2 citations)

4 protocols using ultracentrifuge xl 100 k

Lentiviral Vector Production in HEK293T Cells

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HEK293 T (CRL-3216) cells were purchased from the Cell Bank of Iranian Pasteur
Institute and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented
with 10% fetal bovine serum (FBS) and incubated at 37°C in a humidified
atmosphere containing 5% CO₂⁴⁶.
HEK293 T cells were transfected with the pCDH-CXCR4 lentiviral vectors (21 µg)
(Wild type or Mutant) and two packaging vectors, psPAX2 (21 µg, gifted by
Tronolab) and pMD2 (10 µg) by calcium phosphate method⁴⁶ for the lentivirus production process. Culture media were replaced by
fresh media containing 10% FBS after 16 h, and then viral particles were
harvested at 48, 72, and 96 h post-transfection. Supernatants were filtered
through 0.45 µm filters (Orange Scientific, Braine-l’Alleud, Belgium), pelleted
by ultracentrifugation (Beckman-Coulter ultracentrifuge XL-100 K, Brea, CA, USA)
(at 28,000 rpm, 4°C for 1 h) and re-suspended in serum-free media, to increase
viral concentration⁴⁶.

Bidkhori H.R., Bahrami A.R., Farshchian M., Heirani-tabasi A., Mirahmadi M., Hasanzadeh H., Ahmadiankia N., Faridhosseini R., Dastpak M., Shabgah A.G, & Matin M.M. (2021). Mesenchymal Stem/Stromal Cells Overexpressing CXCR4R334X Revealed Enhanced Migration: A Lesson Learned from the Pathogenesis of WHIM Syndrome. Cell Transplantation, 30, 09636897211054498.

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Lentiviral Particle Production Protocol

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For lentivirus production, HEK293T cells were transfected with pLKO.1/puro plasmids together with pCMV-dR8.91 and pCMV-VSV-G packing plasmids using Calcium Phosphate Transfection Kits (Clontech). Viral particles were collected 48 h after transfection, filtered with 0.45 µm sterile filter and concentrated by ultracentrifugation at 4 °C (24,000 rpm, 2 h, Beckman-Coulter ultracentrifuge XL-100K).

Luo M., Wu L., Zhang K., Wang H., Zhang T., Gutierrez L., O’Connell D., Zhang P., Li Y., Gao T., Ren W, & Yang Y. (2018). miR-137 regulates ferroptosis by targeting glutamine transporter SLC1A5 in melanoma. Cell Death and Differentiation, 25(8), 1457-1472.

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Isolation and Characterization of Endometrial Cell-Derived Exosomes

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Extracellular vesicles released by endometrial ECC-1 cells were isolated via the differential centrifugation of the conditioned media (secretome) [36 (link)]. In brief, five 175 cm² cell culture flasks (Corning, USA) were seeded per cell line and cells were grown until 90% confluence at 37ºC and under 5% CO₂. To avoid exogenous exosome contamination, cells were washed with PBS, incubated for 1 h in FBS-free DMEM, washed with PBS, and incubated with 20 mL of DMEM without FBS for 48 h at 37ºC under 5% CO₂. Then, 100 mL of conditioned medium per cell line were centrifuged at 500 g for 5 min at 4ºC to remove the cell debris (pellet) and then centrifuged again at 2000 g for 10 min at 4ºC to remove vesicles greater than 1 μm (pellet). Next, exosomes were purified by differential centrifugation (Beckman-Coulter ultracentrifuge, XL-100 K, USA) [36 (link)]. Exosomes were analyzed by a NanoSight NS300 (Malvern Panalytical, United Kingdom) and by transmission electron microscopy using a FEI Tecnai 12 electron microscope to verify the quality of the purified exosomes, as previously described [36 (link)]. Exosomes were stored at -80ºC until use.

Montero-Calle A., López-Janeiro Á., Mendes M.L., Perez-Hernandez D., Echevarría I., Ruz-Caracuel I., Heredia-Soto V., Mendiola M., Hardisson D., Argüeso P., Peláez-García A., Guzman-Aranguez A, & Barderas R. (2023). In-depth quantitative proteomics analysis revealed C1GALT1 depletion in ECC-1 cells mimics an aggressive endometrial cancer phenotype observed in cancer patients with low C1GALT1 expression. Cellular Oncology (Dordrecht), 46(3), 697-715.

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Lentivirus Production and Concentration

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For lentivirus production, HEK293T cells were transfected with pLKO.1/puro plasmids together with pCMV-dR8.91 and pCMV-VSV-G packing plasmids using Calcium Phosphate Transfection Kits (Clontech). Viral particles were collected 48 h after transfection, filtered with 0.45 µm sterile filter, and concentrated by ultracentrifugation at 4 °C (24,000 rpm, 2 h, Beckman-Coulter ultracentrifuge XL- 100 K).

Zhang Y., Luo M., Cui X., O’Connell D, & Yang Y. (2022). Long noncoding RNA NEAT1 promotes ferroptosis by modulating the miR-362-3p/MIOX axis as a ceRNA. Cell Death and Differentiation, 29(9), 1850-1863.

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