Millex hv filter

Manufactured by Merck Group

Sourced in United States, Germany

The Millex-HV filter is a sterile, single-use filter device designed for the filtration of aqueous solutions. It is constructed with a hydrophilic polyvinylidene fluoride (PVDF) membrane and has a pore size of 0.45 micrometers. The filter is intended to remove particulates and microbial contaminants from solutions prior to use in laboratory applications.

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Lab products found in correlation

Sep pak c18 plus short cartridge, by Waters Corporation (2 mentions) Millex hpf hv filter, by Merck Group (2 mentions) Sep pak plus light cartridge, by Waters Corporation (2 mentions) Alpha 1 4 lsc freeze dryer, by Martin Christ (2 mentions) Plenti puro vector, by Addgene (2 mentions) Entrez gene id 967, by GenScript (2 mentions) G1322a quaternary pump, by Agilent Technologies (2 mentions) Dulbecco s modified eagle s medium nutrient mixture f 12, by Thermo Fisher Scientific (1 mentions) Medium 199, by Thermo Fisher Scientific (1 mentions) Complete dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Pqcxip vector, by Takara Bio (1 mentions) Pretrox tet3g, by Takara Bio (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) Agilent 6410 triplequad lc ms system, by Agilent Technologies (1 mentions) Lc 6a, by Shimadzu (1 mentions) 4d nucleofector, by Lonza (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Lc 10, by Shimadzu (1 mentions) Discovery c18, by Merck Group (1 mentions) Jurkat, by American Type Culture Collection (1 mentions)

31 protocols using millex hv filter

Propagation and Titration of Tiger Frog Virus

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FHM cells were cultured in Medium 199 (Gibco, Life Technologies, USA) supplemented with 10 % fetal bovine serum (FBS) (Biochrom, Merck Millipore, Darmstadt, Germany) at 27 °C. ZF4 cells were cultured in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 (Gibco, Life Technologies, USA) supplemented with 10 % FBS at 27 °C. HepG2 cells were cultured in complete Dulbecco’s modified Eagle’s medium (Gibco, Life Technologies, USA) supplemented with 10 % FBS at 37 °C in a 2.5 % CO₂-humidified chamber [27 (link)].
TFV was originally isolated from diseased tiger frog tadpoles from Nanhai (Guangdong, China) and is being maintained in our laboratory [22 (link)]. FHM cells were used as the host cells to propagate TFV at 27 °C, and the virus was harvested 3 days after the cytopathic effect was marked. Virus suspension was produced by freeze–thaw the virus–cell mixture three times, followed by filtration with a 0.45 μm Millex-HV Filter (Merck Millipore, Darmstadt, Germany). The virus titer was determined through the 50 % endpoint method using the 50 % tissue culture infective dose (TCID₅₀), and a multiplicity of infection value of 10 was used for infection [28 , 29 (link)].

Yuan J.M., Chen Y.S., He J., Weng S.P., Guo C.J, & He J.G. (2016). Identification and differential expression analysis of MicroRNAs encoded by Tiger Frog Virus in cross-species infection in vitro. Virology Journal, 13, 73.

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Retroviral transduction of murine eIF3e

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HA‐tagged murine eIF3e, eIF3e‐ΔC 31, and AcGFP were subcloned into the NotI‐BamHI site of the pQCXIP vector (TAKARA, Shiga, Japan) and were cotransfected with into GP2‐293 cells along with pVSV‐G to produce viral particles. Retroviral supernatants were harvested 48 h after transfection and concentrated by PEG centrifugation. After filtration by Millex‐HV filter (Merck Millipore), these retroviral supernatants were used to transduce genes into MEFs.

Sadato D., Ono T., Gotoh‐Saito S., Kajiwara N., Nomura N., Ukaji M., Yang L., Sakimura K., Tajima Y., Oboki K, & Shibasaki F. (2018). Eukaryotic translation initiation factor 3 (eIF3) subunit e is essential for embryonic development and cell proliferation. FEBS Open Bio, 8(8), 1188-1201.

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Generation and Cultivation of Cell Lines for HIV-1 Research

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HEK293, HEK293T (ATCC) and TZM-bl (NIH AIDS Reagent Program) cells were maintained in DMEM supplemented with 10% fetal bovine serum. CEM, CEMSS, H9, M8166 (NIH AIDS Reagent Program) and human PBMCs (purchased from Kurabo, Osaka, Japan) were cultured in RPMI containing 10% fetal bovine serum. To generate CEM-TetON-ASK1 or CEMSS-TetON-ASK1 cells, parental cells were co-infected with VSV-G-pseudotyped retroviral vectors expressing pRetroX-TRE3G-ASK1 and pRetroX-Tet3G (Clontech) and selected with G418 and puromycin. HIV-1 stocks were produced by transient transfection of HEK293T cells with the pNL4-3 (ref. 56 (link)) or AZT-resistant pNLGRINFQ molecular clones (NIH AIDS Reagent Program). Culture supernatants containing virus were collected at 48 h after transfection, filtered through a 0.45-μm Millex-HV filter (Merck Millipore, Billerica, MA) and immediately stored until use.

Miyakawa K., Matsunaga S., Kanou K., Matsuzawa A., Morishita R., Kudoh A., Shindo K., Yokoyama M., Sato H., Kimura H., Tamura T., Yamamoto N., Ichijo H., Takaori-Kondo A, & Ryo A. (2015). ASK1 restores the antiviral activity of APOBEC3G by disrupting HIV-1 Vif-mediated counteraction. Nature Communications, 6, 6945.

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Standardized Extraction of Chinese Herbs

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The concentrated Chinese medicine granules (CCMG) of Hedyotis diffusa (HD) and Artemisia capillaris (AC) (Guangdong Yifang Pharmaceutical Co., Ltd., Foshan, China) are crude aqueous extracts from 15 g raw herb plants. Both CCMGs were dissolved in 15 mL of EMEM supplemented with heat-inactivated 10% FCS, then passed through a sterile 0.45 µm membrane filtration (Millex-HV filter, Merck Millipore Ltd., Tullagreen, Ireland) to make stock solutions, which equated to 1000 mg raw herbs per mL (1000 mg/mL). The concentrate was further aliquoted into smaller sizes and stored at -20°C until further experiments. All stored herbs were used once after thaw.

Mao Z.Q., Minakawa N, & Moi M.L. (2022). Novel Antiviral Efficacy of Hedyotis diffusa and Artemisia capillaris Extracts against Dengue Virus, Japanese Encephalitis Virus, and Zika Virus Infection and Immunoregulatory Cytokine Signatures. Plants, 11(19), 2589.

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Quantification of Flavonoids in Scutellaria baicalensis

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The quantification of baicalin, baicalein, wogonoside, and wogonin was carried out using an Agilent 6410 Triple Quad LC/MS system. Root samples of S. baicalensis were collected and homogenized to obtain a fine powder, which was then subjected to ultrasonic extraction with 70% (v/v) ethanol at 60°C for 1 h. The obtained extracts were centrifuged at 10,000 g at 4°C for 15 min and subsequently filtered through a Millipore Millex-HV filter (Merck Millipore, Billerica, MA, USA). To determine the flavone content, an absolute quantification method with external standards was employed. Calibration curves were constructed using standard solutions of baicalin, baicalein, wogonoside, and wogonin at known concentrations. The analytical parameters of the Agilent 6410 Triple Quad LC/MS were as follows: column, Waters T3 column (2.1 mm × 100 mm, 3 μm); oven temperature, 350°C. The mobile phase contained D: water (containing 0.1% [v/v] formic acid); C: acetonitrile; flow rate, 0.3 mL/min; column temperature, 50°C; and injection volume, 5 μL.

Fang S., Zhang C., Qiu S., Xiao Y., Chen K., Lv Z, & Chen W. (2023). SbWRKY75- and SbWRKY41-mediated jasmonic acid signaling regulates baicalin biosynthesis. Frontiers in Plant Science, 14, 1213662.

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Essential Amino Acid Profile of P. pacifica

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The essential amino acid profile of P. pacifica was determined by examining its essential amino acid contents. Essential amino acid analysis was conducted using highperformance liquid chromatography (HPLC) type 1100 system with a Eurospher 100-5 C18, 250 × 4.6 mm column with a P/N: 1115Y535 pre-column. The effluents were: A) 0.01 M acetate buffer at pH 5.9; and B) 0.01 M MeOH acetate buffer at pH 5.9; THF> 80:15:5 Λ Fluorescence: Ext: 340 mm Em: 450 nm. About 2.5 g of sample was put into a sealed glass and 15 mL 6M HCl was added. Subsequently, the mixture was vortexed for homogeneity and subjected to hydrolysis in an autoclave at 110ºC for 12 h, before being cooled to room temperature and neutralized with 6M NaOH. After the addition of 2.5 mL 40% lead acetate and 1 mL 15% oxalate acid, approximately 3 mL of the mixture was filtered with a 0.45 µm Millex-HV filter (Merck KGaA, Darmstadt, Germany). Twenty-five microliters of the filtered mixture plus 475 µL of OPA anhydrolase solution were vortexed and incubated for 3 min for injection into the HPLC system. Finally, 30 µL of the final mixture was placed into the HPLC system. The amino acid composition of the sample was determined using a HPLC system (Shimadzu LC-6A, Shimadzu, Kyoto, Japan) (AOAC 2005; Herawati et al. 2017) (link).

Herawati V.E., Pinandoyo P., Ariati R.W., Rismaningsih N., Windarto S., Prayitno S.B., Darmanto Y., & Radjasa O.K. (2021). Effects of Caulerpa lentillifera added into culture media on the growth and nutritional values of Phronima pacifica, a natural fish-feed crustacean. Biodiversitas Journal of Biological Diversity, 22(1).

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HIV-1 production and CEM cell infection

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Replication-competent HIV-1 stocks were produced by transient transfection of HEK293 cells with the pNL4-3 or pNL4-3DNef plasmids. Culture supernatants containing HIV-1were collected 48 h after transfection and filtered through a 0.45 µm Millex-HV filter (Merck).
CEM cells (10 6 cells) were transfected with 50 pmol of siRNAs using a 4D-Nucleofector (Lonza) with electroporation program CL-120, following the manufacturer's instructions. At 24 h posttransfection, the CEM cells were infected with 25 ng of the HIV-1 p24 antigen. The cells were then centrifuged for 90 min at 800 x g with 5 µg/ml polybrene, and subsequently washed three times with PBS to remove the added viruses. Thereafter, we periodically collected the nascent virions that were produced by the infected cells and measured the p24 levels as described in section 2.3.

Miyakawa K., Nishi M., Jeremiah S.S., Morikawa Y., & Ryo A. (2022). MAL Inhibits the Production of HIV-1 Particles by Sequestering Gag to Intracellular Endosomal Compartments. Frontiers in Virology, 2.

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Quantification of Dihydrocapsaicin in Ferulago stellata

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The dried 100 mg of the ethanol and water extracts of kınkor (Ferulago stellata) were dissolved in water in a 5 mL volumetric flask, which was kept in an ultrasonic bath until a clear solution was obtained. Then, 0.1 mL of dihydrocapsaicin solution, used as an internal standard, was added and diluted to the volume with mobile phase and stirred and heated to get clear solution. Then, the solution was filtered (0.45 µm Millipore Millex-HV filter). The concentration of final solution (1 mL) was added in a capped auto sampler vial, from which 2 µL of sample was injected to LC for each run. The prepared samples in the auto sampler were stored at 15 °C [99 ,100 ,101 (link)].

Kızıltaş H., Bingol Z., Gören A.C., Kose L.P., Durmaz L., Topal F., Alwasel S.H, & Gulcin İ. (2021). LC-HRMS Profiling and Antidiabetic, Anticholinergic, and Antioxidant Activities of Aerial Parts of Kınkor (Ferulago stellata). Molecules, 26(9), 2469.

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Activating ASI1-ER Fusion Protein in U2OS Cells

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The BF strain of human breast fibroblasts [54] (link) and the Hs68 strain of human foreskin fibroblasts (ATCC: CRL 1635) were propagated as previously described [55] (link). U2OS cells expressing AsiSI:ER and a pBABE-based retroviral vector encoding the HA-tagged AsiSI:ER fusion protein were generously provided by Dr Gaëlle Legube [26] (link). To generate infectious viral particles, the vector was transfected into 293 T cells expressing the structural components for amphotropic retroviruses. Medium was harvested after 24–36 h, filtered through a 0.45 µm Millex-HV filter (Millipore) and used directly. The recipient cells were passaged (1∶4) into 100 mm culture dishes 24 h prior to infection. The medium was replaced with 10 ml of the filtered viral supernatant. After 48 h, the cells were washed and placed in selection medium containing 0.75 µg/ml puromycin (Invitrogen).
To activate the AsiSI:ER fusion protein, cells were grown to near confluence and incubated in medium containing 4-hyrodoxy tamoxifen (OHT) at a final concentration of 300 nM. Control cells received an equivalent amount of methanol, which was used as a solvent for the OHT. For most experiments, cells were fixed after 4 h and analyzed by either immunofluorescence or chromatin immunoprecipitation (ChIP).

Chandler H., Patel H., Palermo R., Brookes S., Matthews N, & Peters G. (2014). Role of Polycomb Group Proteins in the DNA Damage Response – A Reassessment. PLoS ONE, 9(7), e102968.

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Solid-Phase Extraction of Biomolecules

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The supernatants were filtered through syringe filters without pre-filter (Millex-HV Filter, 0.45 µm, PVDF, Millipore) or syringe filters with a graduated glass fibre pre-filter (Millex-HPF HV Filter, 0.45 μm, PVDF, Millipore). The filtrates of the biological fluids were acidified by trifluoroacetic acid (TFA) (final concentration 0.1% vol/vol). In the SPE technology, depending on the type and size of the sample, Sep-Pak Plus Light Cartridge (130 mg of C18 sorbent per cartridge, Waters, Milford, MA), or Sep-Pak C18 Plus Short Cartridge (360 mg of C18 sorbent per cartridge, Waters, Milford, were used according to the producer’s protocol using a Baker Vacuum Manifold SPE-12G unit (J.T.Baker, Germany). The volume of eluate was reduced on a miVac centrifugal vacuum concentrator, model DNA-23050-800 with SpeedTrap (Genevac Limited, UK) for two hours. Then samples were lyophilized using an ALPHA 1-4 LSC freeze dryer (MARTIN CHRIST Gefriertrocknungsanlagen GmbH Germany) and stored at − 80 °C until analysis.
The chemicals used for extraction: glacial acetic acid (cat. no. 951503, J.T. Baker), trifluoroacetic acid –TFA (cat. no. 9470, J.T. Baker) and acetonitrile-LC–MS reagent (cat. no. 9821.1000, J.T. Baker) were of high purity grade—HPLC grade.

Mikołajczyk A, & Złotkowska D. (2019). Subclinical lipopolysaccharide from Salmonella Enteritidis induces neuropeptide dysregulation in the spinal cord and the dorsal root ganglia. BMC Neuroscience, 20, 18.

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