Transwell kit

Manufactured by BD

Sourced in United States

The Transwell kit is a cell culture insert system designed for the study of cell migration and permeability. The kit consists of a membrane-containing insert that sits within a companion well, allowing for the creation of an upper and lower chamber. This setup enables the monitoring of cell movement and the passage of substances across the membrane.

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Lab products found in correlation

Cell counting kit 8 cck 8 kit, by Dojindo Laboratories (1 mentions) Growth factor reduced biocoat matrigel invasion chambers, by BD (1 mentions) Doxycycline, by Merck Group (1 mentions) Antibody against β actin, by Merck Group (1 mentions) Phosphor erk, by Cell Signaling Technology (1 mentions) Sdf 1α, by Merck Group (1 mentions) Xfect transfection agent, by Takara Bio (1 mentions) Total erk, by Cell Signaling Technology (1 mentions) Mmp 2, by Cell Signaling Technology (1 mentions) Mmp 9, by Cell Signaling Technology (1 mentions) Cisplatin, by Merck Group (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Transwell assay kit Transwell cell migration assay kit

8 protocols using transwell kit

Cell Proliferation and Migration Assay

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Cell proliferation analysis was conducted with a Cell Counting Kit-8 (CCK-8) kit (Dojindo, Kumamoto, Japan) according to the manufacturer’s instructions. After establishing the K562-shRNA model, cells were divided into three groups for further study, including shRNA-γ-catenin cells (knockdown, K562/KD), shRNA-NC cells (transfection negative control, K562/NC), and normal cells (K562 without transfection, K562/WT). In every group, the cell viability was determined at 0, 12, 24, 36, 48, and 60 hours posttransfection using the CCK-8 reagent. Each assay was run in triplicate and repeated three times.

Relative cell viability (%) = \frac{{OD}_{450} of K 562 / KD or K 562 / NC}{{OD}_{450} of K 562 / WT}

Inhibitor rate = (1 - {OD}_{450} - \frac{Treated}{{OD}_{450}} - Untreated) \times 100 %

Cell migration experiment was conducted with the Transwell kit (Becton, Dickinson and Company) according to the instructions of manufacturer.

Xu J., Wu W., Shen W, & Liu P. (2016). The clinical significance of γ-catenin in acute myeloid leukemia. OncoTargets and therapy, 9, 3861-3871.

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Transwell Migration and Invasion Assay

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Migration and invasion assays for PC-3, DU-145, PZ-HPV-7, and C4-2B cells were performed with a transwell kit from Becton Dickinson (Franklin Lakes, NJ, USA). PC-3, DU-145, and C4-2B cells with or without ROR2 overexpression or RWPE-1 cells with or without ROR2 shRNA knockdown were seeded (2 × 10⁴) into DMEM without serum at upper compartment of transwell. The lower compartment of transwell was loaded with DMEM containing 10% FBS. The cell migration and invasion chambers were inserted into the lower compartment and incubated for 24 h. Cells migrated to another side of filter were fixed with cold methanol for 10 min, and then stained with Hematoxylin solution (0.02%) for 30 min. Cells remained on the topside of the filter were removed with a cotton swab. Migration and invasiveness were evaluated by counting the invading cells under a light microscope. All experiments were conducted in triplicates.

Tseng J.C., Huang S.H., Lin C.Y., Wang B.J., Huang S.F., Shen Y.Y, & Chuu C.P. (2020). ROR2 suppresses metastasis of prostate cancer via regulation of miR-199a-5p–PIAS3–AKT2 signaling axis. Cell Death & Disease, 11(5), 376.

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DU-145 Cell Migration Assay

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Migration assays with DU-145 cells were performed using a Transwell kit from BD Biosciences (Franklin Lakes, NJ, USA). Cells were removed from culture plates with trypsin and washed twice with PBS. Cells (4×10⁴) in 250 µl Dulbecco's modified Eagle's medium (DMEM) with or without 0.1 and 2 mg/ml GLT were placed in the upper invasion chamber and the lower compartment was loaded with DMEM supplemented with 10% FBS. The cell migration chamber was inserted into the lower compartment and incubated for 24 h at 37°C. Cells on the upper side of the filter were removed with a cotton swab. Cells attached to the filter were fixed with 100% methanol for 10 min at room temperature. Cells attached to the filter were stained with Giemsa stain (5%) for 1 h at room temperature. Filters were destained by washing with water and the number of cells attached to the filter was then quantified by enumerating cells in images captured using a light microscope of the stained filters.

Qu L., Li S., Zhuo Y., Chen J., Qin X, & Guo G. (2017). Anticancer effect of triterpenes from Ganoderma lucidum in human prostate cancer cells. Oncology Letters, 14(6), 7467-7472.

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Transwell Invasion Assay for Cancer Cells

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Transwell kit (8.0μm pore size polycarbonate filter) with a Matrigel overlay (BD, NJ) was used to evaluate the invasion ability of cells. 786-O cells and ACHN cells (1×10⁵) with 200μL of FBS-free medium was added into the upper chamber, and 600μL of medium with 20% FBS was add into the lower chamber. After incubating for 24 hours at 37°C with 5% CO2, non-invasive cells were removed on the upper surface by a cotton swab. The invaded cells were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet and counted.

Chen Z., Zhang Y., Wu X., Zhang J., Xu W., Shen C, & Zheng B. (2021). Gαi1 Promoted Proliferation, Migration and Invasion via Activating the Akt-mTOR/Erk-MAPK Signaling Pathway in Renal Cell Carcinoma. OncoTargets and therapy, 14, 2941-2952.

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Transwell Migration and Invasion Assays for Prostate Cancer Cells

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Migration assays for control, AR siRNA knockdown, or Casodex‐treated (5 μmol/L Casodex treatment for three passages) C4‐2B cells were carried out following the instructions of the Transwell kit purchased from BD Bioscience (Franklin Lakes, NJ, USA) as previously described.31 Cells were seeded at a concentration of 2 × 10⁴ in 500 μL serum‐free RPMI medium into the upper compartment of Transwell. The lower compartment of Transwell was filled with 500 μL RPMI medium containing 10% FBS. After 18 hours incubation, cells that migrated to the lower surface of the filter were fixed with 100% methanol and stained with 1% Giemsa solution. Total migrated cells were counted following standard procedures. Invasion assay of control or AR siRNA knockdown C4‐2B cells was carried out with Growth Factor Reduced BD BioCoat Matrigel invasion chambers according to the manufacturer's instructions (BD Bioscience) as previously described.31 Cells were seeded at a concentration of 2 × 10⁴ in 500 μL serum‐free RPMI medium into the upper compartment of Transwell. The lower compartment of Transwell was filled with 500 μL RPMI medium containing 10% FBS. After 18 hours incubation, cells that migrated to the lower surface of the filter were fixed with 100% methanol and stained with 1% Giemsa solution. Total migrated cells were counted following standard procedures.

Lin C., Jan Y., Kuo L., Wang B., Huo C., Jiang S.S., Chen S., Kuo Y., Chang C, & Chuu C. (2018). Elevation of androgen receptor promotes prostate cancer metastasis by induction of epithelial‐mesenchymal transition and reduction of KAT5. Cancer Science, 109(11), 3564-3574.

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Ovarian Cancer Cell Line Characterization

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Human ovarian cancer cell lines HO8910 was obtained from department of pathophysiology of Second Military Medical University. SKOV3 and its cognate Cisplatin-resistant cells SKOV3/DDP were obtained from ATCC (American Tissue Culture Collection). SKOV3.ip cells were provided by the Department of Gynecology and Obstetrics, Shanghai First People's Hospital (obtained from ATCC). Cells were maintained in RPMI-1640 medium with 10% fetal bovine serum at 37°C in a humidified 5% CO₂ atmosphere. Cisplatin, doxycycline, SDF-1α, antibody against β-actin were purchased from Sigma-Aldrich. Antibodies against CXCR4, phosphor-Akt, total Akt, phosphor-ERK, total ERK, MMP-2, MMP-9, caspase-3 were purchased from Cell Signaling Technology. Transwell kit was purchased from BD, USA. ELISA kit was purchased from R&D, USA. Cell apoptotic kit was purchased from Roche, USA. Small interfering RNAs were synthesized by Genepharma Company (Shanghai, China). Xfect Transfection Agent was purchased from Clontech, USA. RNeasy minikit was purchased from QIAGEN, Clifton Hill, Australia.

Wu W., Yu L.H., Ma B, & Xu M.J. (2014). The Inhibitory Effect of Doxycycline on Cisplatin-Sensitive and -Resistant Epithelial Ovarian Cancer. PLoS ONE, 9(3), e89841.

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Transwell Assay for PC-3 and DU-145 Cell Migration

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Migration assays for PC-3 and DU-145 cells were performed following the instructions of the transwell kit purchased from BD Bioscience (Franklin Lakes, NJ, U.S.A.) and as previously described [44 (link)]. PC-3 and DU-145 cells seeded at a concentration of 2 × 10⁴/250 μl into upper compartment of transwell. The lower compartment of transwell was filled with 800 μl complete medium. PC-3 and DU-145 PCa cells were pre-treated with 0, 20, 40 and 80 μM CAPE for 24 h. After appropriate incubation time, total number of migrated cells was counted following standard procedures. Cells were then removed from tissue culture plates with trypsin, and washed once with DMEM serum free medium.

Tseng J.C., Lin C.Y., Su L.C., Fu H.H., Yang S.D, & Chuu C.P. (2016). CAPE suppresses migration and invasion of prostate cancer cells via activation of non-canonical Wnt signaling. Oncotarget, 7(25), 38010-38024.

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Transwell Assay for Cell Migration

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Transwell assays were performed using a Transwell kit (BD Bioscience, CA, USA). Briefly, HepG2 cells (8×10⁴ cells) were suspended in 300 μl of serum-free DMEM medium and placed in the upper chamber of the insert; complete medium was added into the lower chamber. After incubation for 36 h, the cells were fixed with methanol and stained with Giemsa; the cells on the top surface of the membrane were wiped off and the cells on the lower surface were analyzed under a microscope (Olympus DP72, Olympus, Tokyo, Japan). The average number of migrated cells was determined to measure the migration capacity.

Xie M.H., Fu Z.L., Hua A.L., Zhou J.F., Chen Q., Li J.B., Yao S., Cai X.J., Ge M., Zhou L, & Wu J. (2022). A new core–shell-type nanoparticle loaded with paclitaxel/norcantharidin and modified with APRPG enhances anti-tumor effects in hepatocellular carcinoma. Frontiers in Oncology, 12, 932156.

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