GATA4 Knockout Sertoli Cell Line
Product classification
Welcome to consult Elem's high-throughput CRISPR service.
Contact us to orderGATA4 Knockout Sertoli Cell Line
Product ID： LM02194021944
Market price
Stock surplus
0
Antibody custom service consulting
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: GATA4 Knockout Sertoli Cell Line
- Commodity ID: LM02194021944
- Gene Symbol: Gata4, Gata-4
- Ensembl ID: ENSMUSG00000021944
- Uniprot ID: Q08369
- 宿主细胞 / 类型: Sertoli/TM4 normal mouse testicular Sertoli cells
- NCBI Gene ID: 14463
- 规格: 1×10^6 cells/frozen vial
- 生长培养基: DMEM+10%FBS+1%P/S
- 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 1/3 to 1/8 confluency
- 倍增时间: ~24h
- 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: Transcriptional activator that binds to the consensus sequence 5'-AGATAG-3' and plays a key role in cardiac development (PubMed:33941892, PubMed:35182466). In cooperation with TBX5, it binds to cardiac super-enhancers and promotes cardiomyocyte gene expression, while it down-regulates endocardial and endothelial gene expression (By similarity). Involved in bone morphogenetic protein (BMP)-mediated induction of cardiac-specific gene expression (By similarity). Binds to BMP response element (BMPRE) DNA sequences within cardiac activating regions (By similarity). Acts as a transcriptional activator of ANF in cooperation with NKX2-5 (PubMed:9584153). Promotes cardiac myocyte enlargement (By similarity). Required during testicular development (By similarity). May play a role in sphingolipid signaling by regulating the expression of sphingosine-1-phosphate degrading enzyme, sphingosine-1-phosphate lyase (By similarity).
- 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.
-
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: Passaged at a ratio of 1/3 to 1/4; cells will confluence 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. - In antibody validation
Previous Page
Next Page
Classification： Gene Knockout Cell Line（Cell Line）
Cell Line Information
Gene Symbol
Gata4, Gata-4
NCBI Gene ID
14463
Ensembl ID
ENSMUSG00000021944
Uniprot ID
Q08369
Screening marker
N/A
Host cell/type
Sertoli/TM4 normal mouse testicular Sertoli cells
Specifications
1×10^6 cells/frozen vial
Growth Medium
DMEM+10%FBS+1%P/S
growth characteristics
Adherent cells, epithelial-like
culture condition
Incubator at 37°C, 5% CO2; passage at 1/3 to 1/8 confluency
doubling time
~24h
Reference fluid change frequency
2-3 times/week
Mycoplasma test results
Negative
Knock-out validation
Knockout efficiency (Sanger sequencing)
100%
Proteome Validation Results
N/A
Antibody number
Adding...
Antibody validation results
Cell Line Description
Introduction of target gene
Transcriptional activator that binds to the consensus sequence 5'-AGATAG-3' and plays a key role in cardiac development (PubMed:33941892, PubMed:35182466). In cooperation with TBX5, it binds to cardiac super-enhancers and promotes cardiomyocyte gene expression, while it down-regulates endocardial and endothelial gene expression (By similarity). Involved in bone morphogenetic protein (BMP)-mediated induction of cardiac-specific gene expression (By similarity). Binds to BMP response element (BMPRE) DNA sequences within cardiac activating regions (By similarity). Acts as a transcriptional activator of ANF in cooperation with NKX2-5 (PubMed:9584153). Promotes cardiac myocyte enlargement (By similarity). Required during testicular development (By similarity). May play a role in sphingolipid signaling by regulating the expression of sphingosine-1-phosphate degrading enzyme, sphingosine-1-phosphate lyase (By similarity).
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: Passaged at a ratio of 1/3 to 1/4; cells will confluence 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.
Related Products
Copyright © Elem Biotechnology (Wuhan) Co., Ltd.
LM