PEI 1 mg mL−1 was batch tested and added at a ratio of 6:1 PEI: DNA. The solution was briefly vortexed and incubated at room temperature for 15 min. Following this the solution was added dropwise to the cells. Flasks were rocked gently in a circular motion to distribute the precipitates, and then returned to the incubator. Twenty-four hours later fresh DMEM supplemented with 10% FBS and 1% penicillin/streptomycin was added. The viral supernatant was collected 72 h after transduction, cleared by centrifugation at 500 × g for 5 min at 4 °C then passed through a 0.45 µm pore PVDF Millex-HV (Millipore). Lentivirus was aliquoted and stored at −80 °C for future use.
Millex hv pvdf
The Millex-HV PVDF is a laboratory filtration product manufactured by Merck Group. It is a high-flow, low-protein-binding syringe filter made from polyvinylidene fluoride (PVDF) material. The core function of the Millex-HV PVDF is to filter samples prior to analysis or further processing.
Lab products found in correlation
11 protocols using millex hv pvdf
Lentiviral Vector Production in HEK293T Cells
PEI 1 mg mL−1 was batch tested and added at a ratio of 6:1 PEI: DNA. The solution was briefly vortexed and incubated at room temperature for 15 min. Following this the solution was added dropwise to the cells. Flasks were rocked gently in a circular motion to distribute the precipitates, and then returned to the incubator. Twenty-four hours later fresh DMEM supplemented with 10% FBS and 1% penicillin/streptomycin was added. The viral supernatant was collected 72 h after transduction, cleared by centrifugation at 500 × g for 5 min at 4 °C then passed through a 0.45 µm pore PVDF Millex-HV (Millipore). Lentivirus was aliquoted and stored at −80 °C for future use.
Ibuprofen Tablets Dissolution Analysis
Lentiviral Particle Generation Protocol
Lentiviral Particle Production Protocol
Lentiviral Particle Production Protocol
Standardized pH Measurement Protocol
Nasal Lining Fluid Proteomic Analysis
HPLC Analysis of Chemical Compounds
filter the sample. Using an analytical column (Phenomenex® ODS 100 A 250 mm ×
4.60 mm, 5 μm) accompanied by a C18 guard column (2.0 cm 4.0 mm; 5 m), HPLC
analysis was carried out on a Shimadzu chromatographer outfitted with a ternary
pump (Shimadzu LC-20AT) and diode assay detector (Shimadzu SPD-M20A, Kyoto,
Japan). Data processing was carried out using LC solutions software (version
1.25). Acetonitrile and water were used as the mobile phase in a gradient
chromatography procedure, with the initial acetonitrile/water ratio being 2:8
v/v and the final acetonitrile/water ratio being 8:2 v/v at 1 mL/min flow rate
over the course of 30 min with 25oC column temperature and 20 μL
injection volume. Every day, the mobile phase was prepared and sonicated to
remove gas before usage. The UV spectra were observed between 450 and 200 nm.
All of the standard solutions and extracts were made with ethanol. The
concentration of the solution chemical was 2000 g/mL, whereas the reference
solutions were employed in 6 different concentrations (12.5, 25.0, 100.0, 150.0,
250.0, and 500.0 g/mL).22
HIV-eGFP Viral Infection Assay
CTR, M1-MDM and M12 MDM were infected with 60 µl/well of HIV-eGFP viral stock. Infected MDM (5 × 105 cells/condition) were detached from plastic adhesion after incubation with Accutase (150 µl/well for 30 min at 37 °C), as described46 (link), spun, and their pellet was resuspended in a fixing solution containing 4% paraformaldehyde (PFA). Flow cytometry for GFP expression was performed using a FACS Calibur instrument (Becton Dickinson Italia, SpA, Milano, Italy), and the results were analyzed with the FlowJo software version 8.4.3 (Tree Star, Ashland, Oregon, USA).
Protein Concentration Determination
= 12.0 on Perkin Elmer (Lambda 25) double beam spectrophotometer attached with peltier temperature programmer (PTP-1). A filtered buffer through a 0.45 μm Millipore Millex-HV PVDF filter was used throughout the study.
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