Siglec-E Knockout RAW264.7 Cell Line

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LM02042030474

Product ID： LM02042030474

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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: Siglec-E Knockout RAW264.7 Cell Line
- Commodity ID: LM02042030474
- Gene Symbol: Siglec12 Siglec5 Siglece Siglecl1
- NCBI Gene ID: 83382
- Ensembl ID: ENSMUSG00000030474
- Uniprot ID: Q91Y57
- 宿主细胞 / 类型: RAW264.7/mouse mononuclear macrophage leukemia cells
- 规格: 1×10^6 cells/ cryovial
- 生长培养基: DMEM + 10% FBS + 1% P,S
- 筛选标记: N/A
- 生长特性: Adherent cells, irregular shape
- 培养条件: 37℃, 5% CO2 incubator, passage 1/6 to 1/8
- 倍增时间: ~12-30 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: A putative adhesion molecule that mediates sialic-acid-dependent binding to cells. The sialic acid recognition site may be masked by cis interactions with sialic acids on the same cell surface. In the immune response, it may act as an inhibitory receptor upon ligand-induced tyrosine phosphorylation by recruiting cytoplasmic phosphatase(s) via their SH2 domain(s), which block signal transduction through dephosphorylation of signaling molecules.
- 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) are 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:
- Siglec12 Siglec5 Siglece Siglecl1
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 biological safety cabinet and wipe the surface with 70% ethanol.
04. Unscrew the cap of 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 1/6 to 1/8, confluent in 2-3 days.
01. When the cell confluence in the culture bottle reaches 80%-90% or more, cell passage can be performed.
02. Take out the culture medium, PBS, trypsin (0.25% Trypsin_EDTA Gibco 25200-056), etc. from the 4℃ refrigerator, place them in a 37℃ water bath, and take them out when the temperature is close to 37℃. After spraying 75% alcohol on the surface of the bottle, 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 a 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 PBS. 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 depending on the actual situation. After about 1-2 minutes, when most of the cells detach, add the corresponding volume of complete culture medium to terminate digestion, and gently blow the cells with a 5mL pipette until all 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 add complete culture medium to the culture bottle. Add 13-15mL to T75, and 5mL to 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 cell passage procedure for specific steps).
04. Add freezing solution to resuspend the cells at 1 mL per tube. After mixing well, aliquot into cryovials.
05. Place the cells in a programmable freezing container and freeze them in a -80℃ freezer.
06. Subsequently, transfer the cells to a liquid nitrogen tank for long-term storage.
Antibody validation

None

Siglec-G Knockout RAW264.7 Cell Line

Gene Knockout Kit Information

Gene Knockout Kit Description

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