Col17A1 Knockout caski Cell Line

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LM02106065618

Product ID： LM02106065618

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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: Col17A1 Knockout caski Cell Line
- Commodity ID: LM02106065618
- Gene Symbol: COL17A1 BP180 BPAG2
- Ensembl ID: ENSG00000065618
- Uniprot ID: Q9UMD9
- 宿主细胞 / 类型: Caski/Human cervical cancer intestinal metastasis cells/Human cervical cancer epithelial cells
- NCBI Gene ID: 1308
- 规格: 1×10^6 cells/ cryovials
- 生长培养基: 1640 + 10% FBS + 1% double antibody
- 筛选标记: N/A
- 生长特性: Adherent cells, epithelial-like
- 培养条件: 37℃, 5% CO2 incubator, passage 1/2 to 1/4
- 倍增时间: ~60-80 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: May play a role in the integrity of hemidesmosomes and the attachment of basal keratinocytes to the underlying basement membrane. The 120 kDa linear IgA disease antigen is an anchoring filament component involved in dermal-epidermal cohesion. It is the target of linear IgA bullous dermatosis autoantibodies.
- 细胞开发路径: Generated stable KO Cell Pool using the CRISPR-RNP method; Sanger sequencing results showed a 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 extensive gene discovery research.
Key words:
- COL17A1 BP180 BPAG2
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 1/2 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 the culture medium, PBS, trypsin (0.25% Trypsin_EDTA Gibco 25200-056), etc., out of 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 biosafety cabinet.

03. Take out the culture bottle to be passaged from the incubator, spray 75% alcohol on the bottle body, and place it in a biosafety cabinet.
04. To avoid disrupting the cells, rinse the cells with PBS along the upper wall of the culture bottle, discard the washing solution, and add 2mL to T25.
05. Add the corresponding volume of trypsin (T75 add 1.5mL, T25 add 0.5mL), and gently shake the bottle to evenly distribute the trypsin on 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 detach, add the corresponding volume of complete culture medium to stop digestion, and gently blow the cells until they are all 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 seeding ratio as needed, and supplement the complete culture medium in the culture bottle; add to 13-15mL for T75, 5mL for T25, and add 1% double antibody.
08. Tighten the bottle cap, gently shake the bottle to mix the cells evenly, and 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 senescence.
03. Digest and centrifuge the cells (refer to the subculture procedure for specific steps).
04. Add freezing solution to resuspend the cells at 1 mL per tube. After mixing well, 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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