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CytoSelect™ 96-well Cell Invasion, Fluorometric

CA 适用: 哺乳动物 Fluorometric Cell Samples, Serum Quantitative Pre-coated
产品编号 ABIN2344867
发货至: 中国
  • 适用
    哺乳动物
    检测方法
    Fluorometric
    应用范围
    Cellular Assay (CA)
    品牌
    CytoSelect™
    样品类型
    Serum, Cell Samples
    Analytical Method
    Quantitative
    产品特性
    CytoSelect™ 96-well Cell Invasion Assay Kit utilizes basement membrane-coated inserts to assay the invasive properties of tumor cells. The kit does not require you to prelabel the cells with Calcein AM or remove non-invaded cells (i.e. cotton swabbing). Any invaded cells are first dissociated from the membrane, then lysed and detected with CyQuant® GR Dye. CytoSelect™ 96-well Cell Invasion Assay Kit provides a robust system for the quantitative determination of cell invasion. The kit contains sufficient reagents for the evaluation of 96 samples.
    组件
    1. 96-well ECM Invasion Plate : One sterile 96-well plate containing ECM-coated inserts (see Figure 1 for components)
    2. 96-well Cell Harvesting Tray : One 96-well tray
    3. Cell Detachment Solution : One 20 mL bottle
    4. 4X Lysis Buffer : One 10 mL bottle
    5. CyQuant® GR Dye : One 75 μL tube
    试剂未包括
    1. Invasive cell lines
    2. Cell culture medium
    3. Serum free medium, such as DMEM containing 0.5 % BSA, 2 mM CaCl2 and 2 mM MgCl2
    4. Cell culture incubator (37 °C, 5 % CO2 atmosphere)
    5. Light microscope
    6. 96-well microtiter plate
    7. Microtiter plate reader 3 Top Plate Cover Middle Invasion Plate Membrane Chamber Bottom Feeder Tray : Components of the 96-well ECM Invasion Plate.
  • 应用备注
    Optimal working dilution should be determined by the investigator.
    说明

    • Fully quantify cell invasion with no manual cell counting
    • Plate inserts are precoated with ECM basement membrane

    板类型
    Pre-coated
    实验流程
    The CytoSelect™ 96-well Cell Invasion Assay Kit contains polycarbonate membrane inserts (8 μm pore size) in a 96-well plate. The upper surface of the insert membrane is coated with a uniform layer of dried basement membrane matrix solution. This basement membrane layer serves as a barrier to discriminate invasive cells from non-invasive cells. Invasive cells are able to degrade the matrix proteins in the layer, and ultimately pass through the pores of the polycarbonate membrane. Finally, the invaded cells are dissociated from the membrane and subsequently detected with CyQuant® GR Dye (Invitrogen).
    实验流程
    1. Under sterile conditions, allow the invasion plate to warm up at room temperature for 10 minutes.
    2. Rehydrate the basement membrane layer of the membrane inserts by adding 100 μL of warm, serum-free media to the inner compartment. Incubate at room temperature for 1 hour.
    3. Prepare a cell suspension containing 0.2-2.0 x 106 cells/mL in serum free media. Agents that inhibit or stimulate cell invasion can be added directly to the cell suspension.
    4. Carefully remove the rehydration medium (step 2) from the inserts without disturbing the basement membrane layer. Note: It will not affect the assay performance if a small amount of rehydration medium is left in the compartment 4
    5. Under sterile conditions, separate the cover and membrane chamber from the feeder tray. Add 150 μL of media containing 10 % fetal bovine serum or desired chemoattractant(s) to the wells of the feeder tray.
    6. Place the membrane chamber back into the feeder tray (containing chemoattractant solution). Ensure no bubbles are trapped under the membrane.
    7. Gently mix the cell suspension from step 3 and add 100 μL to the membrane chamber.
    8. Finally, cover the plate and transfer to a cell culture incubator for 12-24 hours.
    9. Just prior to the end of the incubation, pipette 150 μL of prewarmed Cell Detachment Solution into wells of the clean, 96-Well Cell Harvesting Tray (provided).
    10. Carefully remove the 96-well Invasion Plate from the incubator. Separate the membrane chamber from the feeder tray.
    11. Remove the cells/media from the top side of the membrane chamber by aspirating or inverting. Place the membrane chamber into the Cell Harvesting Tray containing 150 μL of Cell Detachment Solution (step 9). Incubate 30 minutes at 37 °C.
    12. Completely dislodge the cells from the underside of the membrane by gently tilting the membrane chamber several times in the Cell Detachment Solution.
    13. Prepare sufficient 4X Lysis Buffer/CyQuant® GR dye solution for all samples by diluting the dye 1:75 in 4X Lysis Buffer (for example, add 5 μL dye to 370 μL of 4X Lysis Buffer).
    14. Add 50 μL of 4X Lysis Buffer/CyQuant® GR dye solution to each well (already containing 150 μL of Cell Detachment Solution). Incubate 20 minutes at room temperature.
    15. Transfer 150 μL of the mixture to a 96-well plate suitable for fluorescence measurement. Read the fluorescence with a fluorescence plate reader at 480 nm/520 nm. 5
    限制
    仅限研究用
  • 储存条件
    4 °C
    储存方法
    Store all components at 4°C.
  • Knappe, Novak, Weina, Bernhardt, Reith, Larribere, Hölzel, Tüting, Gebhardt, Umansky, Utikal: "Directed de-differentiation using partial reprogramming induces invasive phenotype in melanoma cells." in: Stem cells (Dayton, Ohio), (2016) (PubMed).

    Hirahata, Osaki, Kanda, Sugimoto, Yoshioka, Kosaka, Takeshita, Fujiwara, Kawai, Ito, Ochiya, Okada: "PAI-1, a target gene of miR-143, regulates invasion and metastasis by upregulating MMP-13 expression of human osteosarcoma." in: Cancer medicine, (2016) (PubMed).

    Saha, Choi, Kim, Dayem, Yang, Kim, Yin, Cho: "KRT19 directly interacts with β-catenin/RAC1 complex to regulate NUMB-dependent NOTCH signaling pathway and breast cancer properties." in: Oncogene, Vol. 36, Issue 3, pp. 332-349, (2016) (PubMed).

    Chueca, Apostolova, Esplugues, García-González, Lanas, Piazuelo: "Proton Pump Inhibitors Display Antitumor Effects in Barrett's Adenocarcinoma Cells." in: Frontiers in pharmacology, Vol. 7, pp. 452, (2016) (PubMed).

    Zhou, Ma, Tong, Chan, Wong, Ngan: "Nanomechanical measurement of adhesion and migration of leukemia cells with phorbol 12-myristate 13-acetate treatment." in: International journal of nanomedicine, Vol. 11, pp. 6533-6545, (2016) (PubMed).

    Adam, Matt, Christian, Hess-Stumpp, Haegebarth, Hofmann, Algire: "SIAH ubiquitin ligases regulate breast cancer cell migration and invasion independent of the oxygen status." in: Cell cycle (Georgetown, Tex.), Vol. 14, Issue 23, pp. 3734-47, (2015) (PubMed).

    Ismail, Kang, Lee, Kim, Hong: "DJ-1 upregulates breast cancer cell invasion by repressing KLF17 expression." in: British journal of cancer, Vol. 110, Issue 5, pp. 1298-306, (2014) (PubMed).

    Ruan, Zhang, Li, Yan, Liu, Yang, Wang, Zhang: "Activation of Toll-like receptor-9 promotes cellular migration via up-regulating MMP-2 expression in oral squamous cell carcinoma." in: PLoS ONE, Vol. 9, Issue 3, pp. e92748, (2014) (PubMed).

    Takeuchi, Seike, Noro, Soeno, Sugano, Zou, Uesaka, Nishijima, Matsumoto, Minegishi, Kubota, Gemma: "Significance of osteopontin in the sensitivity of malignant pleural mesothelioma to pemetrexed." in: International journal of oncology, Vol. 44, Issue 6, pp. 1886-94, (2014) (PubMed).

    Yamamoto, Seike, Takeuchi, Soeno, Miyanaga, Noro, Minegishi, Kubota, Gemma: "MiR-379/411 cluster regulates IL-18 and contributes to drug resistance in malignant pleural mesothelioma." in: Oncology reports, Vol. 32, Issue 6, pp. 2365-72, (2014) (PubMed).

    Ichijo, Furuya, Shimura, Hayashi, Takahashi, Ohta, Kobayashi, Kitamura: "Activation of the RhoB signaling pathway by thyroid hormone receptor ? in thyroid cancer cells." in: PLoS ONE, Vol. 9, Issue 12, pp. e116252, (2014) (PubMed).

    Leung, Vong, Lin, Janke, Chen, Leung: "Modulation of NKG2D ligand expression and metastasis in tumors by spironolactone via RXR? activation." in: The Journal of experimental medicine, Vol. 210, Issue 12, pp. 2675-92, (2013) (PubMed).

    Nakayama: "cAMP-response element-binding protein (CREB) and NF-?B transcription factors are activated during prolonged hypoxia and cooperatively regulate the induction of matrix metalloproteinase MMP1." in: The Journal of biological chemistry, Vol. 288, Issue 31, pp. 22584-95, (2013) (PubMed).

    Desai, Reed, Burks, Wood, Pullikuth, Haas, Liu, Breslin, Meiners, Sankar: "ISG15 disrupts cytoskeletal architecture and promotes motility in human breast cancer cells." in: Experimental biology and medicine (Maywood, N.J.), Vol. 237, Issue 1, pp. 38-49, (2012) (PubMed).

    Chen, Zhang, Yue, Zheng, Kovacevic, Richardson: "The iron chelators Dp44mT and DFO inhibit TGF-?-induced epithelial-mesenchymal transition via up-regulation of N-Myc downstream-regulated gene 1 (NDRG1)." in: The Journal of biological chemistry, Vol. 287, Issue 21, pp. 17016-28, (2012) (PubMed).

    Beach, Hussey, Miller, Chaudhury, Patel, Monslow, Zheng, Keri, Reizes, Bresnick, Howe, Egelhoff: "Myosin II isoform switching mediates invasiveness after TGF-?-induced epithelial-mesenchymal transition." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, Issue 44, pp. 17991-6, (2011) (PubMed).

    Lam, Chiu, Chung, Lee, Lee, Koistinen, Koistinen, Seppala, Ho, Yeung: "Glycodelin-A as a modulator of trophoblast invasion." in: Human reproduction (Oxford, England), Vol. 24, Issue 9, pp. 2093-103, (2009) (PubMed).

    Eckstein, Servan, Hildebrandt, Pölitz, von Jonquières, Wolf-Kümmeth, Napierski, Hamacher, Kassack, Budczies, Beier, Dietel, Royer-Pokora, Denkert, Royer: "Hyperactivation of the insulin-like growth factor receptor I signaling pathway is an essential event for cisplatin resistance of ovarian cancer cells." in: Cancer research, Vol. 69, Issue 7, pp. 2996-3003, (2009) (PubMed).

    Neil, Johnson, Nemenoff, Schiemann: "Cox-2 inactivates Smad signaling and enhances EMT stimulated by TGF-beta through a PGE2-dependent mechanisms." in: Carcinogenesis, Vol. 29, Issue 11, pp. 2227-35, (2008) (PubMed).

    Thal, Xavier, Rosentreter, Linder, Friedrichs, Waha, Pietsch, Stumpf, Noegel, Clemen: "Expression of coronin-3 (coronin-1C) in diffuse gliomas is related to malignancy." in: The Journal of pathology, Vol. 214, Issue 4, pp. 415-24, (2008) (PubMed).

  • 背景
    The ability of malignant tumor cells to invade normal surrounding tissue contributes in large part to the significant morbidity and mortality of cancers. Invasiveness requires several distinct cellular functions including adhesion, motility, detachment, and extracellular matrix proteolysis. Metastatic cells produce many proteolytic enzymes (e.g. lysosomal hydrolysates, collagenases, plasminogen activators) while the expression of certain cell surface protease receptors is also increased.
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