Slc25a11 Knockout aml12 Cell Line
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Product ID： LM02113014606
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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
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- Commodity name: Slc25a11 Knockout aml12 Cell Line
- Commodity ID: LM02113014606
- Gene Symbol: Slc25a11, Ogc
- Ensembl ID: ENSMUSG00000014606
- Uniprot ID: Q9CR62
- 宿主细胞 / 类型: Aml12/mouse normal hepatocytes
- NCBI Gene ID: 67863
- 规格: 1×10^6 cells/frozen vial
- 生长培养基: Specialized culture medium
- 筛选标记: N/A
- 生长特性: Adherent cells, epithelial-like
- 培养条件: Incubator at 37°C, 5% CO2; passage at 1/4 to 1/8 confluency
- 倍增时间: ~24-48 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: Protein-level validation has been completed.
- 抗体货号: Adding...
- 目标基因介绍: Catalyzes the transport of 2-oxoglutarate (alpha-oxoglutarate) across the inner mitochondrial membrane in an electroneutral exchange for malate (By similarity). Can also exchange 2-oxoglutarate for other dicarboxylic acids such as malonate, succinate, maleate and oxaloacetate, although with lower affinity (By similarity). Substrate exchange across the membrane occurs consecutively with one substrate being transported first, then dissociating from the substrate binding site before the second substrate binds for transport in the opposite direction (By similarity). Does not transport glutathione (By similarity). In addition can facilitate proton transport in the presence of protonophores such as long-chain fatty acids, contributing to mitochondrial uncoupling and regulation of mitochondrial energetic efficiency (PubMed:38577966). Contributes to several metabolic processes, including the malate-aspartate shuttle, the oxoglutarate/isocitrate shuttle, gluconeogenesis from lactate, and nitrogen metabolism (By similarity). Maintains mitochondrial fusion and fission events, and the organization and morphology of cristae (By similarity). Involved in the regulation of apoptosis (PubMed:21448454).
- 细胞开发路径: A stable KO cell line was generated using the CRISPR-RNP method; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.
- 应用: Gene knockout cell lines (KO Cell Lines) with high knockout efficiency are particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad gene discovery studies.
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01. Pre-warm the complete culture medium in a 37°C water bath.
02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
03. Transfer the cryovial to a biosafety cabinet and wipe its surface with 70% ethanol.
04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
05. Centrifuge at 125g for 5–7 minutes at room temperature, then discard the supernatant.
06. Resuspend the cell pellet in 5 mL of complete culture medium and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
08. Recommended passage ratio: Passage at a dilution of 1/4 to 1/8; cultures become confluent in 2–3 days. -
01. Cell passage can be performed when the cell confluence in the culture flask reaches 80%–90% or higher.
02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin-EDTA Gibco 25200-056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out, spray the exterior of the bottles with 75% ethanol, and then transfer them to a biosafety cabinet.03. Remove the culture flask to be passaged from the incubator, spray the flask with 75% ethanol, and place it in a biosafety cabinet.
04. To avoid detaching the cells, rinse them with PBS along the inner wall of the culture flask; discard the wash solution, and add 2 mL of PBS to a T25 flask.
05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask; 0.5 mL for a T25), and gently swirl the flask so the enzyme covers the bottom. Adjust the volume as needed. After approximately 1–2 minutes, when most cells have detached, add the corresponding volume of complete culture medium to stop the digestion, and use a 5-mL pipette to gently pipette up and down until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300g for 5 min, and discard the supernatant.
07. Resuspend the cells in 5 mL of complete medium, adjust the seeding ratio as needed, and replenish the complete medium in the culture flasks—add up to 13–15 mL for T75 flasks and 5 mL for T25 flasks—and supplement with 1% double antibiotic.
08. After capping the bottle and tightening the cap, gently shake the bottle to mix the cells evenly, then place it in a 37°C, 5% CO2 incubator. -
01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements; examine the following conditions under a microscope: healthy appearance and morphological characteristics, appropriate growth phase (late logarithmic phase), and absence of contamination or signs of decline.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation solution at 1 mL per tube, mix thoroughly by pipetting, and aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a -80°C freezer.
06. The cells are subsequently transferred to a liquid nitrogen tank for long-term storage. - Western blot (WB) analysis was performed simultaneously on lysates from wild-type and KO cells using a specific monoclonal antibody; the target protein band was detected in WT cells but absent or diminished in KO cells.
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Classification： Gene-knockout cell line (protein levels validated)
Cell Line Information
Gene Symbol
Slc25a11, Ogc
NCBI Gene ID
67863
Ensembl ID
ENSMUSG00000014606
Uniprot ID
Q9CR62
Screening marker
N/A
Host cell/type
Aml12/mouse normal hepatocytes
Specifications
1×10^6 cells/frozen vial
Growth Medium
Specialized culture medium
growth characteristics
Adherent cells, epithelial-like
culture condition
Incubator at 37°C, 5% CO2; passage at 1/4 to 1/8 confluency
doubling time
~24-48 hours
Reference fluid change frequency
2-3 times/week
Mycoplasma test results
Negative
Knock-out validation
Knockout efficiency (Sanger sequencing)
100%
Proteome Validation Results
Protein-level validation has been completed.
Antibody number
Adding...
Antibody validation results
Cell Line Description
Introduction of target gene
Catalyzes the transport of 2-oxoglutarate (alpha-oxoglutarate) across the inner mitochondrial membrane in an electroneutral exchange for malate (By similarity). Can also exchange 2-oxoglutarate for other dicarboxylic acids such as malonate, succinate, maleate and oxaloacetate, although with lower affinity (By similarity). Substrate exchange across the membrane occurs consecutively with one substrate being transported first, then dissociating from the substrate binding site before the second substrate binds for transport in the opposite direction (By similarity). Does not transport glutathione (By similarity). In addition can facilitate proton transport in the presence of protonophores such as long-chain fatty acids, contributing to mitochondrial uncoupling and regulation of mitochondrial energetic efficiency (PubMed:38577966). Contributes to several metabolic processes, including the malate-aspartate shuttle, the oxoglutarate/isocitrate shuttle, gluconeogenesis from lactate, and nitrogen metabolism (By similarity). Maintains mitochondrial fusion and fission events, and the organization and morphology of cristae (By similarity). Involved in the regulation of apoptosis (PubMed:21448454).
Cell development path
A stable KO cell line was generated using the CRISPR-RNP method; Sanger sequencing confirmed 100% knockout efficiency in the KO cell line.
Application
Gene knockout cell lines (KO Cell Lines) with high knockout efficiency are particularly well suited for preliminary functional analyses, the development of complex disease models, precision drug screening, and broad gene discovery studies.
Cell Culture Instructions
Cell Resuscitation
01. Pre-warm the complete culture medium in a 37°C water bath.
02. Thaw the cryovial in a 37°C water bath for 1–2 minutes.
03. Transfer the cryovial to a biosafety cabinet and wipe its surface with 70% ethanol.
04. Unscrew the cap of the cryovial and gently transfer the cell suspension into a sterile centrifuge tube containing 9 mL of complete culture medium.
05. Centrifuge at 125g for 5–7 minutes at room temperature, then discard the supernatant.
06. Resuspend the cell pellet in 5 mL of complete culture medium and transfer the cell suspension to a T25 culture flask.
07. Transfer the cells to a 37°C incubator with 5% CO2 for culture.
08. Recommended passage ratio: Passage at a dilution of 1/4 to 1/8; cultures become confluent in 2–3 days.
cell passage
01. Cell passage can be performed when the cell confluence in the culture flask reaches 80%–90% or higher.
02. Remove the culture medium, PBS, and trypsin (0.25% Trypsin-EDTA Gibco 25200-056) from the 4°C refrigerator, place them in a 37°C water bath, and once their temperature approaches 37°C, take them out, spray the exterior of the bottles with 75% ethanol, and then transfer them to a biosafety cabinet.
03. Remove the culture flask to be passaged from the incubator, spray the flask with 75% ethanol, and place it in a biosafety cabinet.
04. To avoid detaching the cells, rinse them with PBS along the inner wall of the culture flask; discard the wash solution, and add 2 mL of PBS to a T25 flask.
05. Add the appropriate volume of trypsin (1.5 mL for a T75 flask; 0.5 mL for a T25), and gently swirl the flask so the enzyme covers the bottom. Adjust the volume as needed. After approximately 1–2 minutes, when most cells have detached, add the corresponding volume of complete culture medium to stop the digestion, and use a 5-mL pipette to gently pipette up and down until all cells are fully detached.
06. Transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 300g for 5 min, and discard the supernatant.
07. Resuspend the cells in 5 mL of complete medium, adjust the seeding ratio as needed, and replenish the complete medium in the culture flasks—add up to 13–15 mL for T75 flasks and 5 mL for T25 flasks—and supplement with 1% double antibiotic.
08. After capping the bottle and tightening the cap, gently shake the bottle to mix the cells evenly, then place it in a 37°C, 5% CO2 incubator.
cell cryopreservation
01. Prepare the cryopreservation solution and pre-cool it in advance.
02. Ensure that the cells to be cryopreserved meet the cryopreservation requirements; examine the following conditions under a microscope: healthy appearance and morphological characteristics, appropriate growth phase (late logarithmic phase), and absence of contamination or signs of decline.
03. Digest and centrifuge the cells (refer to the subculture protocol for specific steps).
04. Resuspend the cells in cryopreservation solution at 1 mL per tube, mix thoroughly by pipetting, and aliquot into cryovials.
05. Place the cells in a programmed cooling box and freeze them in a -80°C freezer.
06. The cells are subsequently transferred to a liquid nitrogen tank for long-term storage.
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