STAMBP Knockout HAP1 Cell pool

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LM01010124356

Product ID： LM01010124356

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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: STAMBP Knockout HAP1 Cell pool
- Commodity ID: LM01010124356
- Gene Symbol: STAMBP AMSH
- NCBI Gene ID: 10617
- Ensembl ID: ENSG00000124356
- Uniprot ID: O95630
- 宿主细胞 / 类型: HAP1/Chronic myeloid leukemia
- 规格: 1×10^6 cells/ cryovials
- 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 ratio 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: Zinc metalloprotease that specifically cleaves 'Lys-63'-linked polyubiquitin chains. It does not cleave 'Lys-48'-linked polyubiquitin chains (By similarity). It plays a role in signal transduction for cell growth and MYC induction mediated by IL-2 and GM-CSF. It potentiates BMP (bone morphogenetic protein) signaling by antagonizing the inhibitory action of SMAD6 and SMAD7. It has a key role in the regulation of cell surface receptor-mediated endocytosis and ubiquitin-dependent sorting of receptors to lysosomes. Endosomal localization is required for efficient EGFR degradation but not for its internalization (By similarity). It is involved in the negative regulation of PI3K-AKT-mTOR and RAS-MAP signaling pathways.
- 细胞开发路径: A stable KO cell pool was generated using the CRISPR-RNP method; Sanger sequencing results showed that the knockout efficiency of the KO cell pool was >70%.
- 应用: Highly efficient gene knockout cell pools (KO Cell Pool) are particularly suitable for preliminary functional analysis, the development of complex disease models, precision drug screening, and a wide range of gene discovery research. KO pools can be directly applied to various types of assays and analyses without the cumbersome process of single-clone selection, significantly improving experimental efficiency.
Key words:
- STAMBP AMSH
01. Prewarm 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%, it 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. Take them out when the temperature is close to 37℃, spray 75% alcohol on the surface of the bottle, and then 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 then place it in the biosafety cabinet.
04. To avoid dispersing 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 (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 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 to 13-15mL for T75, and 5mL for T25, and add 1% double antibody.
08. After tightening the bottle cap, gently shake the bottle to mix the cells evenly, and then place them in a 37℃, 5% CO2 incubator.
01. Prepare freezing solution and pre-chill.
02. Ensure 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 1mL per tube. After mixing well, dispense into freezing tubes.
05. Place the cells in a programmable freezing container and freeze in a -80℃ refrigerator.
06. Subsequently, transfer the cells to a liquid nitrogen tank for long-term storage.
Antibody validation

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