CytoSelect™ 24-well Collagen Cell Invasion Assay, Fluorometric
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Quick Overview for CytoSelect™ 24-well Collagen Cell Invasion Assay, Fluorometric (ABIN2344865)
适用
检测方法
应用范围
样品类型
Analytical Method
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产品特性
- CytoSelect™ Collagen Cell Invasion Assay Kit utilizes Bovine Type I Collagen-coated inserts to assay the invasive properties of tumor cells. It contains sufficient reagents for the evaluation of 12 samples.
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组件
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- Collagen Invasion Chamber Plate : One 24-well plate containing 12 collagen-coated cell culture inserts
- Cell Detachment Solution : One 5 mL bottle
- 4X Lysis Buffer : One 5 mL bottle
- CyQuant® GR Dye : One 25 μL tube
- Forceps : One each
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试剂未包括
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- Invasive cell lines
- Cell culture medium
- Serum free medium, such as DMEM containing 0.5 % BSA, 2 mM CaCl2 and 2 mM MgCl2
- Cell culture incubator (37 °C, 5 % CO2 atmosphere)
- Light microscope
- 96-well plate suitable for a fluorescence plate reader
- Fluorescence plate reader
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Interleukin 1, beta (IL1B) ELISA KitIL1B 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 小鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 人 Colorimetric Sandwich ELISA 62.5 pg/mL - 4000 pg/mL Plasma, Serum
Amyloid beta 1-40 (Abeta 1-40) ELISA KitAbeta 1-40 适用: 人 Colorimetric Sandwich ELISA 7.813 pg/mL - 500 pg/mL Plasma, Serum, Tissue Homogenate
Amyloid beta 1-42 (Abeta 1-42) ELISA KitAbeta 1-42 适用: 人 Colorimetric Sandwich ELISA 4.688 pg/mL - 300 pg/mL Plasma, Serum, Tissue Homogenate
Lipopolysaccharides (LPS) ELISA Kit适用: Various Species Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 小鼠 Colorimetric Sandwich ELISA 46.88 pg/mL - 3000 pg/mL Plasma, Serum
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 人 AA 77-233 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
Interleukin 6 (IL6) ELISA KitIL6 适用: 人 Colorimetric Sandwich ELISA 7.8-500 pg/mL Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate, Urine
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应用备注
- Optimal working dilution should be determined by the investigator.
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说明
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- Fully quantify cell invasion with no manual cell counting
- Plate inserts are precoated with Collagen I gel layer
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板类型
- Pre-coated
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实验流程
- The CytoSelect™ Collagen Cell Invasion Assay Kit contains polycarbonate membrane inserts (8 μm pore size) in a 24-well plate. The upper surface of the insert membrane is coated with a uniform layer of dried Bovine Type I Collagen matrix. This collagen matrix layer serves as a barrier to discriminate invasive cells from non-invasive cells. Invasive cells are able to degrade the collagen matrix layer, and ultimately pass through the pores of the polycarbonate membrane. Finally, these invaded cells are then dissociated from the membrane and subsequently detected by the patented CyQuant® GR Dye (Invitrogen).
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实验流程
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- Under sterile conditions, allow the collagen invasion chamber plate to warm up at room temperature for 10 minutes.
- Rehydrate the collagen layer of the cell culture inserts by adding 300 μL of warm, serum-free media to the inner compartment. Incubate at room temperature for 30 minutes.
- Prepare a cell suspension containing 0.5-1.0 x 106 cells/mL in serum free media. Agents that inhibit or stimulate cell invasion can be added directly to the cell suspension. Note: Overnight starvation may be performed prior to running the assay
- Carefully remove 250 μL of rehydration medium (step 2) from the inserts without disturbing the collagen layer (leaving 50 μL inside).
- Add 250 μL of the cell suspension solution to the inside of each insert.
- Add 500 μL of media containing 10 % fetal bovine serum or desired chemoattractant(s) to the lower well of the invasion plate.
- Incubate for 12-24 hours in a cell culture incubator.
- Carefully aspirate the media from the inside of the insert. Transfer the insert to a clean well containing 225 μL of Cell Detachment Solution. Incubate 30 minutes at 37 °C.
- Completely dislodge the invaded cells from the underside of the membrane by gently tilting the insert several times in the detachment solution. Remove and discard the insert.
- 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 to 370 μL of 4X Lysis Buffer).
- Add 75 μL of 4X Lysis Buffer/CyQuant® GR dye solution to each well containing cells and 225 μL of Cell Detachment Solution. Incubate 20 minutes at room temperature.
- Transfer 200 μL of the mixture to a 96-well plate suitable for fluorescence measurement. Read fluorescence with a fluorescence plate reader at 480 nm/520 nm. 4
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限制
- 仅限研究用
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储存条件
- 4 °C
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储存方法
- Store all components at 4°C.
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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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