MDK Knockout LLC1 Cell Line

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LM02018027239

Product ID： LM02018027239

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隐藏域元素占位

产品描述
细胞复苏
细胞传代
细胞冻存
抗体验证结果

- Commodity name: MDK Knockout LLC1 Cell Line
- Commodity ID: LM02018027239
- Gene Symbol: Mdk Mk
- NCBI Gene ID: 17242
- Ensembl ID: ENSMUSG00000027239
- Uniprot ID: P12025
- 宿主细胞 / 类型: LLC1/mouse lung cancer cells
- 规格: 1×10^6 cells/ cryovials
- 生长培养基: DMEM + 10% FBS + 1% P,S
- 筛选标记: N/A
- 生长特性: Semi-attached, epithelial-like
- 培养条件: 37℃, 5% CO2 incubator, passage 1/2 to 1/4
- 倍增时间: ~24-36 hours
- 参考换液频率: 2~3 times/week
- 支原体检测结果: Negative
- 敲除效率（Sanger测序）: 100%
- 蛋白质组验证结果: N/A
- 抗体货号: Adding
- 目标基因介绍: Secreted protein that functions as a cytokine and growth factor and mediates its signal through cell-surface proteoglycan and non-proteoglycan receptors. Binds cell-surface proteoglycan receptors via their chondroitin sulfate (CS) groups. Thereby regulates many processes like inflammatory response, cell proliferation, cell adhesion, cell growth, cell survival, tissue regeneration, cell differentiation and cell migration. Participates in inflammatory processes by exerting two different activities. Firstly, mediates neutrophils and macrophages recruitment to the sites of inflammation both by direct action by cooperating namely with ITGB2 via LRP1 and by inducing chemokine expression.
- 细胞开发路径: A stable KO Cell Pool was generated using the CRISPR-RNP method; Sanger sequencing results showed 100% knockout efficiency in the KO Cell Pool
- 应用: Highly efficient gene knockout cell lines (KO Cell Line), particularly suitable for preliminary functional analysis, the development of complex disease models, precision drug screening, and a wide range of gene discovery research.
Key words:
- Mdk Mk
01. Pre-warm the complete medium in a 37℃ water bath.
02. Thaw the cryovials in a 37℃ water bath for 1-2 minutes.
03. Transfer the cryovials to a biosafety cabinet and wipe the surface with 70% ethanol.
04. Unscrew the cryovials and gently transfer the cell suspension to a sterile centrifuge tube containing 9mL of complete medium.
05. Centrifuge at 125g for 5-7 minutes at room temperature and discard the supernatant.
06. Resuspend the cell pellet with 5mL of complete medium and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37℃, 5% CO2 incubator for culture.
08. Reference passage ratio: passage 1/2 to 1/4, confluent in 2-3 days.
01. When the cell confluence in the culture bottle reaches 80%-90%, the cells can be passaged.
02. Take out the culture medium, PBS, trypsin (0.25%Trypsin_EDTA Gibco 25200-056), etc., from the 4℃ refrigerator and place them in a 37℃ water bath. When the temperature is close to 37℃, take them out, spray 75% alcohol on the surface of the bottle, and place them in a biological safety cabinet.

03. Take out the culture bottle to be passaged from the incubator, spray 75% alcohol on the bottle, and place it in a biological safety cabinet.
04. To avoid disrupting the cells, rinse the cells with PBS along the upper wall of the culture bottle. After washing the cells, discard the solution. Add 2mL to T25.
05. Add the corresponding volume of trypsin (1.5mL for T75, 0.5mL for T25), and gently shake the bottle to spread the trypsin evenly over the bottom of the cells. The amount can be appropriately increased or decreased according to the actual situation. After about 1-2 minutes, when most of the cells have detached, add the corresponding volume of complete culture medium to terminate digestion, and gently blow the cells until all cells have detached using a 5mL pipette.
06. Transfer the cell suspension to a 15mL centrifuge tube, centrifuge at 300g for 5min, and discard the supernatant.
07. Resuspend the cells with 5mL of complete culture medium, adjust the inoculation ratio as needed, supplement the complete culture medium in the culture bottle, add 13-15mL to T75, 5mL to T25, and add 1% double antibody.
08. Cover the bottle cap tightly, gently shake the bottle to mix the cells evenly, and place it in a 37℃, 5% CO2 incubator.
01. Prepare freezing solution and pre-chill.
02. Ensure that the cells to be frozen meet the freezing requirements. Check the following status under a microscope: healthy appearance and morphological characteristics, growth cycle (late logarithmic phase), no signs of contamination or degeneration.
03. Digest and centrifuge the cells (refer to the subculture procedure for specific steps)
04. Add freezing solution to resuspend cells at 1 mL per tube, mix well, and dispense into freezing tubes.
05. Place the cells in a programmable freezing container and freeze in a -80℃ freezer.
06. Subsequently transfer the cells to a liquid nitrogen tank for long-term storage.
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