0.45 μm pore size filter

Manufactured by Sartorius

Sourced in Germany

The 0.45-μm pore size filter is a laboratory filtration device designed to remove particles and contaminants from liquid samples. It features a pore size of 0.45 micrometers, which is suitable for a variety of filtration applications. The filter can be used to clarify and purify solutions, remove suspended solids, and prepare samples for further analysis or processing.

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Lab products found in correlation

Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) M17 broth, by Thermo Fisher Scientific (1 mentions) 0.22 μm pore filter, by Sartorius (1 mentions) Agencourt ampure xp system, by Beckman Coulter (1 mentions) Qubit 2.0 fluorometer, by Thermo Fisher Scientific (1 mentions) Polyethylenimine (pei), by Polysciences (1 mentions)

6 protocols using 0.45 μm pore size filter

Propagation and Characterization of Dairy Phages

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Streptococcus thermophilus and Lactococcus lactis strains and phages used for this study are listed in Table 5. Host strains were stored at −40°C in growth medium supplemented with 15% (wt/vol) glycerol. S. thermophilus strains were cultured overnight at 37°C in LM17 broth (M17 broth [Oxoid, Denmark] with 2% [wt/vol] lactose). L. lactis strains were cultured overnight at 30°C in GM17 broth (M17 broth [Oxoid] with 0.5% [wt/vol] glucose).
Phages were propagated by transferring a single plaque into 10 ml of LM17- or GM17-Ca/Mg broth (i.e., growth medium supplemented with 10 mM CaCl₂ and 10 mM MgCl₂) inoculated with 1% overnight culture of adequate host and incubating until full lysis had occurred. The volume was gradually brought to 20 ml by adding host culture growing in LM17- or GM17-Ca/Mg. Upon lysis, the pure phage lysates were filtered through 0.45-μm-pore-size filters (Sartorius, Germany). The propagated phages were stored at 4°C.
Phage titers as well as the host ranges of investigated phages with selected bacterial strains were determined by using the double agar overlay spot test, as described previously (27 (link)). Following overnight incubation under the appropriate growth conditions, the PFU per milliliter were calculated.

Szymczak P., Janzen T., Neves A.R., Kot W., Hansen L.H., Lametsch R., Neve H., Franz C.M, & Vogensen F.K. (2017). Novel Variants of Streptococcus thermophilus Bacteriophages Are Indicative of Genetic Recombination among Phages from Different Bacterial Species. Applied and Environmental Microbiology, 83(5), e02748-16.

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Stress Resistance Assays for Microbial Strains

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Stress resistance assays were performed as in de Dios et al. (2020 (link)). Briefly, to test the resistance to osmotic stress and copper, 10 μl spots of serial dilutions of late‐exponential phase cultures were placed on solid MML rich medium plates supplemented with NaCl 0.6 M or CuSO₄ 3.5 mM and incubated for 5 days at 30°C. For desiccation assays, 5 μl spots of serial dilutions of late‐exponential phase cultures were placed on 0.45 μm pore size filters (Sartorius Stedim Biotech GmbH) and they were left to air‐dry in a laminar flow cabin for 5 h (5 min in the control assay). Then, filters were placed on MML rich medium plates supplemented with bromophenol blue 0.002% and incubated for 5 days at 30°C. In the case of recovery from oxidative shock, late‐exponential phase cultures were diluted to an OD₆₀₀ of 0.1 in MML medium. When an OD₆₀₀ 0.5 was reached, H₂O₂ was added to the medium in a final concentration of 10 mM. Recovery from the treatment is represented by a percentage of the OD₆₀₀ reached by treated cultures after 5 h of growth compared to non‐treated cultures. At least three independent replicates of each experiment were performed, and most representative examples are shown.

de Dios R., Santero E, & Reyes‐Ramírez F. (2022). The functional differences between paralogous regulators define the control of the general stress response in Sphingopyxis granuli TFA. Environmental Microbiology, 24(4), 1918-1931.

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Green Synthesis of Elderflower-Derived Gold Nanoparticles

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ES stem was purchased from Kwang Myoung Herb Medicine (Busan, South Korea) in September 2016. To prepare ES extract, chopped ES stem was extracted in distilled water at 100°C for 4 hrs. ES supernatant was vacuum-filtered (0.45-μm pore size filter; Sartorius Biotech GmbH, Gottingen, Germany), freeze-dried (final yield of 27 g), and stored in a refrigerator (4°C). ES freeze-dried extract was dissolved in distilled water and filter-sterilized through a 0.22-μm pore filter (Sartorius Biotech GmbH, Gottingen, Germany) and stored in a refrigerator (4°C). For the green synthesis of ES-GNs, a mixture of a 1 mM chloroauric acid solution and ES extract (2 mg/mL) was added under rigorous stirring (the mixture immediately turned violet) and was stationarily incubated at room temperature (22–25°C) for 10 mins. Then, the remaining residues were eliminated by centrifugation at 13,000 rpm for 20 mins. ES-GNs were synthesized using the abovementioned procedure but without the addition of chloroauric acid.

Park S.Y., Yi E.H., Kim Y, & Park G. (2019). Anti-neuroinflammatory effects of Ephedra sinica Stapf extract-capped gold nanoparticles in microglia. International Journal of Nanomedicine, 14, 2861-2877.

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Screening Antibacterial Activity of LAB Isolates

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The initial screening of fish isolates was carried out against pathogenic bacteria using agar well diffusion assay (Murray et al., 1995 , Rather et al., 2013 (link)). Briefly, the plates of nutrient agar were prepared and allowed to solidify. The plates were spread with 200 μl of target bacteria (10⁷ cfu/ml) and allowed to dry for 10 min. The 24 h culture broths of 32 LAB isolates grown in MRS media at 37 °C were centrifuged at 10,000g for 10 min. The supernatant was collected and filter-sterilized through a 0·45-μm-pore-size filter (Sartorius Stedim Biotech, Goettingen, Germany). An autoclaved borer was used to make uniform wells poured with 100 μl filter sterilized cell-free-supernatant of the isolated bacterium. The plates were incubated for 24 h at 37 °C. After incubation, the antibacterial activity was determined by measuring the zones of inhibition against tested bacteria. The assay was done in triplicates.

Paray B.A., Rather I.A., Al-Sadoon M.K, & Fanar Hamad A.S. (2018). Pharmaceutical significance of Leuconostoc mesenteroides KS-TN11 isolated from Nile Tilapia, Oreochromis niloticus. Saudi Pharmaceutical Journal : SPJ, 26(4), 509-514.

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Metagenome Extraction from Stream Sediments

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A mixture of water and sediment samples was taken from the middle of SK stream (Figure 1A), and the sample was maintained at 4°C for less than a week before analysis. The mixture was shaken vigorously prior to metagenome extraction. A 100-mL sample was centrifuged at 1000 × g for 5 min to remove coarse particles, and the water was filtered using a 0.45-μm pore size filter (Sartorius, Göettingen, Germany). The filter membrane was then sliced and subjected to metagenome DNA extraction using the Metagenomic DNA Isolation Kit (Epicentre, Wisconsin, USA), according to the manufacturer's suggested protocol. To increase the purity of the metagenome library, humic acids or other PCR inhibitors were removed using the Agencourt AMPure XP System (Beckman Coulter, Brea, CA, USA). The cleaned metagenome was evaluated by 1% w/v agarose gel electrophoresis, a Nanodrop™ 1000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA), and a Qubit® 2.0 Fluorometer (Invitrogen, Merelbeke, Belgium). Metagenomes extracted from the same sampling site were subsequently analyzed by 16S rRNA sequencing and shotgun metagenome analyses.

Chan C.S., Chan K.G., Tay Y.L., Chua Y.H, & Goh K.M. (2015). Diversity of thermophiles in a Malaysian hot spring determined using 16S rRNA and shotgun metagenome sequencing. Frontiers in Microbiology, 6, 177.

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Viral Vector Production in HEK 293T Cells

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Viral vectors were produced by HEK 293T cells after triple transfection with the transfer plasmid described earlier, a second-generation packaging plasmid lacking vif, vpr, vpu, and nef genes (pCMVΔR8.91) and an envelope plasmid encoding vesicular stomatitis virus G (VSV-G) protein. HEK 293T cells were seeded in 10-cm diameter cell-culture dishes at 5 × 10⁶ cells per plate in DMEM supplemented with 10% FCS. After 24 h, 20 μg of transfer plasmid, 10 μg of packaging construct, and 5 μg of envelope plasmid were diluted in 700 μl of 150 mM NaCl. 700 μl of polyethylenimine solution (PEI, Polysciences) was added slowly to the DNA mixture. DNA-PEI mix was incubated for 5 min at room temperature and then the DNA-PEI complex was added dropwise to the HEK 293T cells in DMEM supplemented with 1% FCS. The cells were incubated at 37°C in a 5% CO₂ humidified atmosphere for 24 h, then the medium was replaced with DMEM with 10% FCS. The supernatant was harvested at day 2 and 3 post-transfection and filtered through a 0.45 μm pore-size filter (Sartorius, Minisart, Göttingen, Germany). The filtered vector particles were concentrated to 1 ml using vivaspin (Vivascience, Bornem, Belgium). Pellets were dissolved in DMEM with 10% FCS, divided in 50 μl aliquot in and stored at −80°C.

Giovannozzi S., Lemmens V., Hendrix J., Gijsbers R, & Schrijvers R. (2020). Live Cell Imaging Demonstrates Multiple Routes Toward a STAT1 Gain-of-Function Phenotype. Frontiers in Immunology, 11, 1114.

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