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Human MDM2 Knockout Cell Line - SK-OV-3

Human MDM2 Knockout Cell Line - SK-OV-3

Cat.No. :  CSC-RT2719

Host Cell:  SK-OV-3 Target Gene:  MDM2

Size:  1x10^6 cells/vial, 1mL Validation:  Sequencing

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2719
Description This cell is a stable cell line with a homozygous knockout of human MDM2 using CRISPR/Cas9.
Target Gene MDM2
Host Cell SK-OV-3
Host Cell Species Homo sapiens (Human)
Size Form 1 vial (>10^6 cell/vial)
Shipping Dry ice package
Storage Liquid nirtogen
Revival Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.
Mycoplasma Negative
Format One frozen vial containing millions of cells
Storage Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.

2. No eating, drinking or smoking while handling the stable line.

3. Wash hands after handling the stable line and before leaving the lab.

4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.

5. All waste should be considered hazardous.

6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship Dry ice
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Background

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The MDM2 oncogene, also known as mouse double differential 2 homolog, is a gene that encodes a protein of the same name. The MDM2 gene was first discovered in transformed mouse cell lines, specifically the 3T3-DM cell line. Mouse studies have shown that overexpression of MDM2 binds to the oncogenic protein Ras, leading to transformation of primary rodent fibroblasts and subsequent tumor formation in nude mice. Subsequently, a human analogue, commonly known as HDM2, was discovered and found to be elevated in various human tumor types, including sarcomas, osteosarcomas, and breast cancer. This finding supports the role of MDM2 as an oncogene that has the potential to initiate cancer when overexpressed or mutated. A key role of MDM2 is as a negative regulator of the p53 tumor suppressor. It does this by acting as an E3 ubiquitin ligase, marking p53 for proteasomal degradation. This interaction is an essential component of a negative feedback loop that is necessary to maintain cellular homeostasis. When p53 is activated, it increases the transcription of MDM2, resulting in elevated levels of MDM2 protein. MDM2 then binds to the N-terminal transcriptional activation domain of p53, inhibiting its transcriptional activity and targeting it for degradation. This action prevents excessive accumulation of p53, thereby avoiding unnecessary cell cycle arrest or apoptosis.
1. Cancer Research: The human MDM2 knockout cell line - SKOV-3 can be used to study cancer biology, particularly ovarian cancer. By eliminating the MDM2 gene, which normally inhibits the tumor suppressor p53, researchers can better understand the role of p53 in cancer progression, treatment resistance, and cellular responses to DNA damage. 2. Drug Development: SKOV-3-MDM2 knockout cells can be used to screen and develop new anticancer drugs. These cell lines provide a unique platform to test the efficacy of drugs targeting the p53 pathway. 3. Genetics Research: This cell line is an important tool for genetic research, especially in studying the effects of MDM2 loss. Researchers can use these cells to analyze downstream genetic pathways and interactions affected by MDM2, providing insights into the genetic vulnerabilities and resilience mechanisms of cancer cells. 4. Functional Genomics: Functional genomics experiments often require the manipulation of key genes to study their roles in various cellular processes. The SKOV-3-MDM2 knockout cell line allows scientists to perform high-throughput screening and functional analysis to evaluate the effects of MDM2 loss on gene expression, protein function, and cell behavior under different conditions. 5. Toxicology and Safety Testing: The human MDM2 knockout cell line - SKOV-3 can be used for toxicology and safety testing of new drugs. By examining the cytotoxic effects of compounds on MDM2-deficient cells, researchers can better evaluate the safety of candidate drugs, especially their interactions with the p53 pathway and potential off-target effects.
Customer Q&As
How is the knockout cell line validated?

A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.

Is the product a single clonal cell or mixed cell pool?

A: Single clonal cell.

Can I confirm gene knockout by RT-qPCR?

A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.

How can I store the cell product?

A: The cell line should be stored in liquid nitrogen for long-term preservation.

Is it possible to get multiple knockout clones for my GOI?

A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.

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Customer Reviews
Versatile Experimental Applications

Whether for drug screening, genetic analysis, or signaling pathway studies, the Human MDM2 Knockout Cell Line-SKOV-3 has delivered outstanding performance every time, making it a staple in our lab's workflow.

French

07/29/2024

Comprehensive customer support

The Creative Biogene's customer support team was incredibly helpful during our setup phase. They provided detailed protocols and troubleshooting tips, ensuring we got the best results from the Human MDM2 Knockout Cell Line-SKOV-3.

French

04/24/2020

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