PLXNB2 Knockout HAP1 Cell pool

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LM01010196576

Product ID： LM01010196576

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产品描述
细胞复苏
细胞传代
细胞冻存
抗体验证结果

- Commodity name: PLXNB2 Knockout HAP1 Cell pool
- Commodity ID: LM01010196576
- Gene Symbol: PLXNB2 KIAA0315
- Ensembl ID: ENSG00000196576
- Uniprot ID: O15031
- 宿主细胞 / 类型: HAP1/Chronic Myelogenous Leukemia
- NCBI Gene ID: 23654
- 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 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 validation
- 抗体货号: Adding
- 目标基因介绍: Cell surface receptor for SEMA4C, SEMA4D, and SEMA4G that plays an important role in cell-cell signaling (By similarity). Plays a role in glutamatergic synapse development and is required for SEMA4A-mediated excitatory synapse development (By similarity). Binding to class 4 semaphorins promotes downstream activation of RHOA and phosphorylation of ERBB2 at 'Tyr-1248' (By similarity). Required for normal differentiation and migration of neuronal cells during brain corticogenesis and for normal embryonic brain development (By similarity). Regulates the migration of cerebellar granule cells in the developing brain (By similarity). Plays a role in RHOA activation and subsequent changes of the actin cytoskeleton (PubMed:12183458). Plays a role in axon guidance, invasive growth, and cell migration (PubMed:15184888). May modulate the activity of RAC1 and CDC42 (By similarity).
- 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, development of complex disease models, precision drug screening, and extensive gene discovery research. KO pools allow for direct application in various assays and analyses without tedious single-clone selection, significantly improving experimental efficiency.
Key words:
- PLXNB2 KIAA0315
01. Pre-warm the complete culture 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 culture 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 culture 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. Take them out when the temperature is close to 37℃, spray 75% alcohol on the surface of the bottle, and then place them in the 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 disrupting the cells, rinse the cells with PBS along the upper wall of the culture bottle. After washing the cells, discard the liquid. 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 evenly distribute the trypsin 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 detach, add the corresponding volume of complete culture medium to terminate the digestion, and gently blow the cells with a 5mL pipette until all the cells detach.
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, 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 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 decline.
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 cryovials.
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.
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