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Cytoselect 24 well cell migration and invasion assay kit

Manufactured by Cell Biolabs
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The CytoSelect 24-well Cell Migration and Invasion Assay kit from Cell Biolabs provides a quantitative measurement of cell migration and invasion through a cell culture system. The kit contains inserts coated with an extracellular matrix, allowing for the assessment of both cell migration and invasion capabilities.

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15 protocols using cytoselect 24 well cell migration and invasion assay kit

1

Cell Cycle, Apoptosis, and Migration Analysis

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FACS analysis for cell cycle and apoptosis was done 72 hours post-transfection. The cells were harvested, washed with cold PBS, and resuspended in the nuclear stain DAPI for cell cycle analysis or stained with 7-AAD and Annexin-V-FITC using ANNEXIN V-FITC/7-AAD KIT (BD Biosciences) for apoptosis analysis according to the manufacturer’s protocol. Stained cells were immediately analyzed by FACS (BD FACSVerse; BD Biosciences). Cell viability was determined at 24, 48 and 72 h by using the CellTiter 96 AQueous One Solution Cell Proliferation Assay kit (Promega, Madison, WI) according to the manufacturer’s protocol. Also a cytoselect 24-well cell migration and invasion assay kit (Cell Biolabs, Inc) was used for migration and invasion assays according to the manufacturer’s protocol. For migration and invasion assays two replicates were used and the experiment was repeated three times.
Bhat N.S., Colden M., Dar A.A., Saini S., Arora P., Shahryari V., Yamamura S., Tanaka Y., Kato T., Majid S, & Dahiya R. (2017). MicroRNA-720 regulates E-cadherin-αE-catenin complex and promotes renal cell carcinoma. Molecular cancer therapeutics, 16(12), 2840-2848.
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2

Cell Migration and Invasion Assay

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Cell migration and invasion were assessed by using CytoSelect 24-well Cell Migration and Invasion Assay kit (cat no. CBA-100-C, Cell Biolabs), according to the manual. In brief, 1.5 × 105 cells in 300 μl of RPMI were placed on an 8-μm pore size polycarbonate membrane insert (for migration) or on a polycarbonate membrane insert coated with a basement membrane layer (invasion assay) in 500 μl of 10% RPMI. After incubation for 24 h (migration) or 48 h (invasion assay), non-migratory cells in the upper chamber were carefully removed with cotton-tipped swabs, and the migratory cells were stained and read by a plate reader (Versamax tunable microplate reader, Molecular devices) at an optical density of 560 nm.
Costa H., Xu X., Overbeek G., Vasaikar S., Patro C.P., Kostopoulou O.N., Jung M., Shafi G., Ananthaseshan S., Tsipras G., Davoudi B., Mohammad A.A., Lam H., Strååt K., Wilhelmi V., Shang M., Tegner J., Tong J.C., Wong K.T., Söderberg-Naucler C, & Yaiw K.C. (2016). Human cytomegalovirus may promote tumour progression by upregulating arginase-2. Oncotarget, 7(30), 47221-47231.
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3

Cell Viability, Invasion, and Migration Assay

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Cell viability was investigated using the Cell Counting Kit 8 (WST-8 / CCK8) (Abcam, USA) according to the manufacturer’s instructions. Cell invasion and migration abilities were determined using the CytoSelect™ 24-Well Cell Migration and Invasion Assay kit (Cell Biolabs Inc, USA) according to the manufacturer’s instructions. Invasive or migratory cells were fixed in methanol and stained with crystal violet (Abcam, USA).
Huang X., Liu X., Du B., Liu X., Xue M., Yan Q., Wang X, & Wang Q. (2021). LncRNA LINC01305 promotes cervical cancer progression through KHSRP and exosome-mediated transfer. Aging (Albany NY), 13(15), 19230-19242.
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4

Assaying Cell Migration and Invasion

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Cell migration and invasion were assayed using a CytoSelect™ 24-Well Cell Migration and Invasion Assay kit (Cell Biolabs, Inc., San Diego, CA) according to the manufacturer’s protocol. In brief, SKOV3 cells were treated with either control or STIP1 siRNA for 48 h, and were subsequently suspended in serum free medium (1.0 × 106 cells/mL). Cell suspension (300 μL) was added to the upper chamber. The membrane chambers were then transferred to the lower well of the migration plate, which was filled with 10% fetal bovine serum-containing media. Cells were allowed to migrate for 24 h at 37°C. Migratory cells that had passed through membrane were collected, lysed, and quantified at OD 560 nm using a plate reader.
For invasion assays, cells were treated with either control or STIP1 siRNA for 48 h, and were subsequently suspended in serum free medium (1.0 × 106 cells/mL). Cell suspension (300 μL) was added to the upper chamber, which was coated with a uniform layer of dried basement membrane matrix solution. The membrane chambers were then transferred to the lower well of the invasion plate, which was filled with 10% fetal bovine serum-containing media. Cells were allowed to invade for 24 h at 37°C. Invasive cells that had passed through the basement membrane layer were collected, lysed, and quantified at OD 560 nm using a plate reader.
Cho H., Kim S., Shin H.Y., Chung E.J., Kitano H., Park J.H., Park L., Chung J.Y., Hewitt S.M, & Kim J.H. (2014). Expression of Stress-Induced Phosphoprotein1 (STIP1) is Associated with Tumor Progression and Poor Prognosis in Epithelial Ovarian Cancer. Genes, chromosomes & cancer, 53(4), 277-288.
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5

Cell Cycle and Apoptosis Analysis

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FACS analysis for cell cycle and apoptosis was done 72 hours (96 hrs for MDAPCab cells) post-transfection using nuclear stain DAPI for cell cycle analysis or ANNEXIN V-FITC /PI KIT (Betkin Dikeson) for apoptosis analysis according to the manufacturer's protocol. Cell viability was determined at 24, 48, 72 and 96 h by using the CellTiter 96 AQueous One Solution Cell Proliferation Assay kit (Promega, Madison, WI) according to the manufacturer's protocol. For colony formation assay, cells were seeded at low density (1000 cells/plate) and allowed to grow until visible colonies appeared. Then, cells were stained with Giemsa and colonies were counted. Cytoselect 24-well cell migration and invasion assay kit (Cell Biolabs, Inc) was used for migration and invasion assays according to manufacturer's protocol.
Nip H., Dar A.A., Saini S., Colden M., Varahram S., Chowdhary H., Yamamura S., Mitsui Y., Tanaka Y., Kato T., Hashimoto Y., Shiina M., Kulkarni P., Dasgupta P., Imai-Sumida M., Tabatabai Z.L., Greene K., Deng G., Dahiya R, & Majid S. (2016). Oncogenic microRNA-4534 regulates PTEN pathway in prostate cancer. Oncotarget, 7(42), 68371-68384.
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6

Assessing Cell Migration and Invasion

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Migration and invasion of BC cells were assessed using CytoSelect 24-well Cell Migration and Invasion Assay kit (CBA-100-C, Cell Biolabs) following manufacturer instructions [27 (link),28 (link)]. In brief, 1.5 × 105 cells placed on an 8-μM pore size insert. After incubation for 24 h or 48 h, the non-migratory/non-invasive cells the upper chamber were carefully removed with cotton-tipped swabs, and the migratory/invasive cells processed per vendor’s protocol and read by a plate reader (Versamax tunable microplate reader, Molecular Devices) at λ560 nm. Before removing the cells from the upper chamber, the non-migratory cells visualized by a Nikon ECLIPSE TE200-U microscope (Nikon Instruments Inc., Melville, NY, USA). Digital images were captured using Nikon NIS Elements software (Nikon Instruments Inc., Melville, NY, USA).
Siddique A.B., Ebrahim H.Y., Akl M.R., Ayoub N.M., Goda A.A., Mohyeldin M.M., Nagumalli S.K., Hananeh W.M., Liu Y.Y., Meyer S.A, & El Sayed K.A. (2019). (−)-Oleocanthal Combined with Lapatinib Treatment Synergized against HER-2 Positive Breast Cancer In Vitro and In Vivo. Nutrients, 11(2), 412.
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7

Cell Migration and Invasion Assay

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The migration and invasion assay was performed using the CytoSelect 24-Well Cell Migration and Invasion Assay kit (8 μm, Colorimetric Format, Cell Biolabs, San Diego, CA), according to the manufacturer's instructions. Briefly, the migration assay plate was incubated at room temperature for 10 min, and an invasion basement membrane layer of the cell culture insert was rehydrated with warm serum-free RPMI 1640 media for 1 h at room temperature. Cells were detached from the culture plate by trypsinization and resuspended in serum-free RPMI 1640 media. RPMI 1640 media containing 10% FBS was added into the lower chamber, and the cell suspensions were applied to the upper chamber of the migration/invasion insert. Chambers were incubated at 37°C for 24 h and the membrane was stained with cell stain solution at room temperature. The stained migratory or invasive cells were analyzed by a Nikon ECLIPS TS100 inverted microscope (Nikon Instruments).
Lee M.S., Lee J., Kim J.H., Kim W.T., Kim W.J., Ahn H, & Park J. (2015). Overexpression of caldesmon is associated with tumor progression in patients with primary non-muscle-invasive bladder cancer. Oncotarget, 6(37), 40370-40384.
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8

Transwell Migration and Invasion Assay

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The transwell migration and invasion assay was performed using the Cytoselect 24-well cell Migration and Invasion Assay kit (Cell Biolabs, San Diego, CA, USA) according to the manufacturer’s instruction.56 (link)
Mi L., Zhu F., Yang X., Lu J., Zheng Y., Zhao Q., Wen X., Lu A., Wang M., Zheng M., Ji J, & Sun J. (2017). The metastatic suppressor NDRG1 inhibits EMT, migration and invasion through interaction and promotion of caveolin-1 ubiquitylation in human colorectal cancer cells. Oncogene, 36(30), 4323-4335.
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9

Cell Migration and Invasion Assay

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Cytoselect 24-well cell migration and invasion assay kits (Cell Biolabs, Inc., San Diego, CA, USA) were used for the migration and invasion assays, according to the manufacturer’s instructions. AGS and MKN45 cell lines transfected with miR-503 mimics or control miR were harvested 72 h after transfection and resuspended in serum-free Opti-minimum essential medium. The cells (10×104 per 500 μl serum-free media) were added to the upper chambers, and the lower chambers were filled with 750 μl media with 10% FBS. The cells were incubated for 16 h at 37°C and 5% CO2 in a tissue culture incubator. After 16 h, the non-migrated/non-invading cells were removed from the upper sides of the Transwell membrane filter inserts with cotton-tipped swabs. Migrated/invaded cells on the lower sides of the inserts were stained, and the absorbance was read at 560 nm according to the manufacturer’s instructions.
PENG Y., LIU Y.M., LI L.C., WANG L.L, & WU X.L. (2014). microRNA-503 inhibits gastric cancer cell growth and epithelial-to-mesenchymal transition. Oncology Letters, 7(4), 1233-1238.
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10

Quantitative Proteomics Using iTRAQ

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The eight-plex iTRAQ kits were acquired from Applied Biosystems (Foster City, CA, USA). Bio-Rad (Hercules, CA, USA) provided all electrophoresis reagents. CytoSelect™ 24-well Cell Migration and Invasion assay kits (8 μm, colorimetric format) were purchased from Cell Biolabs (San Diego, CA, USA). Opti-MEM was acquired from Gibco (San Diego, CA, USA). SRI specific siRNA (HSS110179, HSS110180, HS110181) and Lipofectamine 2000 were purchased from Invitrogen (Carlsbad, CA, USA). Sorcin, CTSZ, Hspbp1 and CSTB antibody were obtained from OriGene (Rockville, MD, USA). The S100A11 antibody used in this study was purchased from Abcam (Cambridge, MA, USA).
Tuo H., Shu F., She S., Yang M., Zou X.Q., Huang J., Hu H.D., Hu P., Ren H., Peng S.F, & Yang Y.X. (2017). Sorcin induces gastric cancer cell migration and invasion contributing to STAT3 activation. Oncotarget, 8(61), 104258-104271.
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