0.4 μm pore

Manufactured by Corning

Sourced in United States

The 0.4-μm pore lab equipment is a filtration device designed for precise separation and purification of materials. It functions by allowing the passage of particles smaller than 0.4 microns in size, while retaining larger particles. This core capability serves a variety of laboratory applications that require high-precision filtration.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Tritc dextran, by Merck Group (1 mentions) Fitc dextran, by Merck Group (1 mentions) Fv3000, by Olympus (1 mentions)

Close spelling variants of this product

Transwell chambers with 0.4-μm pores 0.4‐μm pore polycarbonate membrane inserts 0.4 μm pore size 0.4 μm pore filters 0.4 μm pore membrane Transwell insert with a 0.4-μm pore size membrane 0.4 μm pore-size permeable supports Transwell 0.4‐μm pore‐size filters Transwell filters with 0.4-μm pore Transwell plates with 0.4-μm pore sized inserts

4 protocols using 0.4 μm pore

Coculture of Tregs and Inflamed Synovial Fibroblasts

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Inflamed SFs were cultured and activated with TNFα (10 ng/ml) stimulation for 24 hours before coculture. For a cell-to-cell contact coculture, iT_regs or nT_regs (T_regs: SFs, 10:1) were added to the activated inflamed SFs directly and cocultured in the presence of anti-CD3/CD28–coated.
beads (T_regs: beads, 1:3) for 3 days. For transwell assays, T_regs were cultured in the insert well of a 24-well transwell plate (0.4-μm pore, Corning) in the presence of anti-CD3/CD28–coated beads (T_regs: beads, 1:3) and while SFs were cultured in the lower chamber separately to coculture for 3 days. nT_regs/pT_regs cultured under T_H17-polarizing condition (the same condition for T_H17 cell differentiated from naïve CD4⁺ T cells) or stimulated with exogenous rmIL-6 (100 ng/ml) for 3 days were set up as a positive control. Foxp3 and IL-17A expression in T_reg subsets were determined by flow cytometry. For in vivo transfer, T_regs after being cocultured with or without CIA-SFs for 3 days were labeled with biotin anti-CD4 antibody and anti-biotin microbeads and then subjected to a positive selection by auto MACS to acquire CD4⁺ T cells. These CD4⁺ T cells were used as T_regs for transfer.

Yang S., Zhang X., Chen J., Dang J., Liang R., Zeng D., Zhang H., Xue Y., Liu Y., Wu W., Zhao J., Wang J., Pan Y., Xu H., Sun B., Huang F., Lu Y., Hsueh W., Olsen N, & Zheng S.G. (2020). Induced, but not natural, regulatory T cells retain phenotype and function following exposure to inflamed synovial fibroblasts. Science Advances, 6(44), eabb0606.

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In vitro Blood-Brain Barrier Permeability Assay

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Primary HBMECs were purchased from Cell Systems (ACBRI 376, Kirkland, WA, USA). HBMECs were grown in Clonetics EGM-2 MV media (CC-3202, Lonza, Walkersville, MD, USA), and only up to eight passages were used for experiments. The in vitro BBB model was established in cell culture inserts as described before66 (link). The transwell PET membranes (0.4-μm pore, 11-mm diameter; Corning, Lowell, MA, USA) were coated with collagen (15 μg ml⁻¹) and fibronectin (30 μg ml⁻¹). HBMECs were seeded onto the membrane at a density of 2.5 × 10⁵ cells per membrane (Fig. 2a). Cultures were maintained at 37 °C in humidified 95% air and 5% CO₂ for 4 days to reach confluence. To assess paracellular permeability, 4.4 kDa TRITC-dextran or 70 kDa FITC-dextran (Sigma-Aldrich) were added into the luminal chamber at a concentration of 2 mg ml⁻¹ in 500 μl media. Fluorescence intensity was measured with a fluorescence reader at 30-min intervals for 1–6 h by removing 30 μl media from the lower (abluminal) chamber. The concentrations of tracers in samples were calculated from a standard curve fitted using known concentrations of tracers. Thirty microlitre fresh media was added after each reading. Paracellular permeability was calculated by measuring the diffusion coefficient of tracers from the luminal to the abluminal chamber11 (link).

Shi Y., Zhang L., Pu H., Mao L., Hu X., Jiang X., Xu N., Stetler R.A., Zhang F., Liu X., Leak R.K., Keep R.F., Ji X, & Chen J. (2016). Rapid endothelial cytoskeletal reorganization enables early blood–brain barrier disruption and long-term ischaemic reperfusion brain injury. Nature Communications, 7, 10523.

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Macrophage-Intestinal Cell Co-culture

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U937 cells were induced into activated macrophages with PMA (10 ng/mL) and IL-4 (10 ng/mL) treatment [20 (link)]. Activated macrophages were seeded in the upper chamber with 0.4 μm pore (Corning Life Sciences, Lowell, MA, USA) and were co-cultured with HT-29 and Caco-2 cells in six-well plates at 37 °C and 5% CO₂.

Yu J., Xu Z., Guo J., Yang K., Zheng J, & Sun X. (2021). Tumor-associated macrophages (TAMs) depend on MMP1 for their cancer-promoting role. Cell Death Discovery, 7, 343.

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Hyaluronic Acid Hydrogel for AB Delivery

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The AB-laden HA hydrogel was prepared by incubating 50 μL of AB solution (2400 μg/mL in PBS) with 50 μL HA (5 mg/mL, Bioregen, China) overnight at 4 °C. The self-cross-linking HA had been approved by the China Food & Drug Administration as a physical barrier for clinical administration (no. 20153641542). To confirm the incorporation of ABs with HA hydrogel, the composite hydrogel was assessed using scanning electron microscopy (SEM) (HITACHI S–3000 N, Japan); Dil-labelled AB-laden HA was fabricated and assessed by confocal laser scanning microscopy (CLSM) (Olympus FV3000, Japan). The release profile of ABs from the composite hydrogel submerged in hyaluronidase (50 ng/mL, diluted in PBS) at 37 °C was examined. Briefly, the 100 μL HA hydrogel containing 120 μg ABs was placed in the upper transwell chamber with 0.4-μm pore (Corning, NY) and 200 μL hyaluronidase was added into the lower chamber. 100 μL leaching solution was collected and replaced by 100 μL hyaluronidase at time points 4 h, 12 h, 24 h, 36 h, 48 h, 72 h, 96 h and 120 h until the sample had degraded. The protein content of released ABs was detected by the BCA protein assay kit (Thermo Fisher Scientific, USA). The release efficiency of ABs was calculated as follows:

Xin L., Wei C., Tong X., Dai Y., Huang D., Chen J., Ma L, & Zhang S. (2021). In situ delivery of apoptotic bodies derived from mesenchymal stem cells via a hyaluronic acid hydrogel: A therapy for intrauterine adhesions. Bioactive Materials, 12, 107-119.

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