CEMIP2 Knockout HAP1 Cell pool

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LM01010135048

Product ID： LM01010135048

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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: CEMIP2 Knockout HAP1 Cell pool
- Commodity ID: LM01010135048
- Gene Symbol: CEMIP2 KIAA1412 TMEM2
- NCBI Gene ID: 23670
- Ensembl ID: ENSG00000135048
- Uniprot ID: Q9UHN6
- 宿主细胞 / 类型: HAP1/Chronic myeloid leukemia
- 规格: 1×10^6 cells/ cryovial
- 生长培养基: IMDM + 10% FBS + 1% P/S
- 筛选标记: N/A
- 生长特性: Adherent cells, epithelial-like
- 培养条件: 37℃, 5% CO2 incubator, passage at a ratio of 1:3 to 1:5
- 倍增时间: ~16 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）: >70%
- 蛋白质组验证结果: Completed protein level verification
- 抗体货号: Adding
- 目标基因介绍: Cell surface hyaluronidase that mediates the initial cleavage of extracellular high-molecular-weight hyaluronan into intermediate-size hyaluronan of approximately 5 kDa fragments (PubMed:28246172). Acts as a regulator of angiogenesis and heart morphogenesis by mediating degradation of extracellular hyaluronan, thereby regulating VEGF signaling (By similarity). Is very specific to hyaluronan; not able to cleave chondroitin sulfate or dermatan sulfate (PubMed:28246172).
- 细胞开发路径: Stable KO Cell Pool was generated using the CRISPR-RNP method; Sanger sequencing results showed that the KO Cell Pool knockout efficiency >70%
- 应用: A highly efficient gene knockout cell pool (KO Cell Pool), especially suitable for preliminary functional analysis, the development of complex disease models, precision drug screening, and extensive gene discovery research. KO pools can be directly applied to various types of assays and analyses without the cumbersome process of single-clone selection, greatly improving experimental efficiency.
Key words:
- CEMIP2 KIAA1412 TMEM2
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 cap 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 1/3 to 1/5, 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 approaches 37℃, take them out, spray 75% alcohol on the surface of the bottle, and then 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 then place it in the biosafety cabinet.
04. To avoid disturbing 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 increased or decreased appropriately 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 with a 5mL pipette until all the cells have detached.
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 seeding ratio as needed, and 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 then place it in a 37℃, 5% CO2 incubator.
01. Prepare the freezing solution and pre-chill it.
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 decay.
03. Digest and centrifuge the cells (refer to the subculture procedure for specific steps).
04. Add 1 mL of freezing solution per tube to resuspend the cells, mix well, and dispense into cryovials.
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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