Cls3421

Manufactured by Corning

Sourced in United States

The CLS3421 is a laboratory equipment product from Corning. It is designed to perform core functions related to sample handling and processing in a research or analytical setting. The detailed specifications and intended use cases of this product are not available for this response.

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Lab products found in correlation

Af3626, by R&D Systems (1 mentions) D1844, by Thermo Fisher Scientific (1 mentions) Cls3428, by Corning (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Mcp 1, by R&D Systems (1 mentions)

5 protocols using cls3421

Neutrophil, Monocyte, T-cell, and Eosinophil Migration Assay

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Bone marrow-derived neutrophils (5×10⁵), monocytes (5×10⁵), T cells (1×10⁶), and eosinophils (5×10⁵ cells) were placed at the upper chamber of 24-transwell membrane plates (Corning; CA-3415, CLS3421 or CA-3422). Migration assays were performed with 5μm pore (neutrophils), 8μm pore (monocytes) or 3μm pore transwell inserts (T cells and eosinophils). Lung homogenate supernatant from normal or metastases-bearing lungs was placed at the bottom chamber. Neutralizing anti-IL-33 antibody (0.2μg/ml AF3626, R&D systems) was added and migrated cells were counted by cell counter.

Shani O., Vorobyov T., Monteran L., Lavie D., Cohen N., Raz Y., Tsarfaty G., Avivi C., Barshack I, & Erez N. (2020). Fibroblast-derived IL-33 facilitates breast cancer metastasis by modifying the immune microenvironment and driving type-2 immunity. Cancer research, 80(23), 5317-5329.

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Measuring Endothelial Permeability in Psoriasis

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Endothelial permeability measurements were performed using a Transwell system with HMEC-1 cells seeded in the upper chamber with a 5.0 µm pore size (CLS3421, Corning, USA) until they reached 90% confluence. Freshly isolated CXCR4^lo and CXCR4^hi neutrophils from healthy controls and psoriasis patients (1×10⁵/well) were added to the upper chamber and co-incubated at 37 °C for 6 h. LDHA-IN-3 (50 μM, MCE) was added to block lactate activity. Transwell inserts without stimulation or with DMSO were used as controls. After 6 h, the culture medium was removed and washed with free-cell medium three times. Next, 50 µl of FITC-labeled dextran (D1844, 2.5 mg/mL, 40 kDa, Invitrogen) was added to the upper chamber as a tracer. After 2 h, 100 µl samples were collected from the lower chamber and fluorescence spectrophotometry was performed with an excitation wavelength of 494 nm and an emission wavelength of 521 nm (spectrofluorometer, Varioskan LUX 3020-265, Thermo Scientific) to measure the permeability of the endothelial monolayer.

Chen J., Bai Y., Xue K., Li Z., Zhu Z., Li Q., Yu C., Li B., Shen S., Qiao P., Li C., Luo Y., Qiao H., Dang E., Yin W., Gudjonsson J.E., Wang G, & Shao S. (2023). CREB1-driven CXCR4hi neutrophils promote skin inflammation in mouse models and human patients. Nature Communications, 14, 5894.

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Cell Migration Across Microporous Membranes

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Cells were seeded at sub-confluent concentrations (10-20 thousand cells/cm²) on polycarbonate Transwell membranes with pore diameters of 3 μm (Corning, CLS3414), 5 μm (Corning, CLS3421), and 8 μm (Corning, CLS3428). Membranes were pre-coated with collagen 1 (50 ug/mL). After seeding cells, the filters were maintained at 37°C and 5% CO₂ for 15 hours. Cells were gently removed from either the top or bottom of the membrane with a cotton swab and immediately fixed with paraformaldehyde. To determine the number of cells per unit area, cells were stained with crystal violet and imaged at multiple locations across the membrane with a 10x objective. Cells were manually counted in 800×800 μm² fields of view (12-30 locations per condition). The percentage of cells that cross the membrane (Fig. 5) was then determined by taking the ratio of the number of cells on the bottom of the membrane and the sum of cells on the filter top and bottom.

Patteson A.E., Pogoda K., Byfield F.J., Mandal K., Ostrowska-Podhorodecka Z., Charrier E.E., Galie P.A., Deptuła P., Bucki R., McCulloch C.A, & Janmey P.A. (2019). Loss of vimentin enhances cell motility through small confining spaces. Small (Weinheim an der Bergstrasse, Germany), 15(50), e1903180.

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Transwell Migration Assay with U0126

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Cells cultured to 90% confluence in T25 flasks (LacZ, tAg, TAg, TAg/tAg) were treated with 10 μM U0126 for 24 h. Afterwards, they were suspended with 0.05% trypsin and seeded into the inner chamber of a transwell (5.0 μm pores, CLS3421, Corning, MA, USA), filled with 200 µL of serum-free DMEM/F12 (Gibco™ 11330032) and the outer chamber filled with 600 µL of complete medium in the inner chamber. After 3 h, the cells were fixed with methanol for 5 min, stained with Liu's stain for 2 min, and then washed twice with PBS. Cells present on the lower surface of the filter were counted in four random microscopic fields (200×), while cells on the upper surface were removed using a cotton swab.

Wang J.W., Li Y.J., Wu H.H., Hsu H.H., Chang M.Y., Wang R.Y, & Tian Y.C. (2023). The essential role of the ERK activation in large T antigen of BK polyomavirus regulated cell migration. Virus Research, 336, 199220.

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Transwell Migration of THP-1 Monocytes

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THP-1 monocytes (10⁶ cells/ml) were stained with Calcein/AM (Invitrogen) for 30 min and then loaded into the upper wells of a 24-well transwell plate (CLS3421, Corning). The lower wells contained either vehicle or MCP-1 (R&D Systems, 150 ng/ml). The cells were incubated for 3 h at 37 °C and 5% CO₂. Transmigrated cells were observed and counted in five separate fields at 10 × magnification under a fluorescence microscope.

Yan J., Zhang Y., Yu H., Zong Y., Wang D., Zheng J., Jin L., Yu X., Liu C., Zhang Y., Jiang F., Zhang R., Fang X., Xu T., Li M., Di J., Lu Y., Ma X., Zhang J., Jia W, & Hu C. (2022). GPSM1 impairs metabolic homeostasis by controlling a pro-inflammatory pathway in macrophages. Nature Communications, 13, 7260.

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