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Human B2M Knockout Cell Line-293T

Human B2M Knockout Cell Line-293T

Cat.No. :  CSC-RT0493

Host Cell:  HEK293T Target Gene:  B2M

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

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Cat. No. CSC-RT0493
Description This KO cell line is a stable cell line with a homozygous knockout of human B2M using CRISPR/Cas9.
Target Gene B2M
Gene ID 567
Genotype B2M (-/-)
Host Cell HEK293T
Host Cell Species Homo sapiens (Human)
Cell Type Epithelial
Size 1x10^6 cells/vial, 1mL
SequencingResult Homozygous: 1 bp insertion in exon 1
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.
Media Type Cells were cultured in DMEM supplemented with 10% fetal bovine serum.
Growth Properties Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6.
Freeze Medium Complete medium supplemented with 10% (v/v) DMSO
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.

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The protein encoded by the β2-microglobulin (B2M) gene is essential for the proper functioning of the immune system. B2M is located on chromosome 15 and is highly conserved across species. Structurally, the B2M protein is a small, non-glycosylated polypeptide consisting of approximately 100 amino acids. It pairs with the α chain of the MHC class I molecule, stabilizing the complex and enabling its transport from the endoplasmic reticulum to the cell surface. This interaction is essential for cellular immune surveillance, enabling the immune system to detect and respond to intracellular pathogens, such as viruses and some forms of bacteria, as well as abnormal cells, such as cancer cells. Mutations or alterations in the B2M gene can have significant clinical implications. For example, loss of B2M expression is frequently observed in certain types of cancer and viral infections, including HIV, and is associated with immune evasion. In addition, elevated levels of β2-microglobulin in the blood may indicate the presence of chronic inflammation or renal dysfunction, as this protein is normally filtered out by the kidneys. The B2M gene has also been implicated in autoimmune diseases. Abnormal B2M expression or function may contribute to the pathogenesis of diseases such as rheumatoid arthritis and systemic lupus erythematosus. These diseases result from the immune system inappropriately targeting the body's own tissues, usually due to abnormal antigen presentation.
The creation of a human B2M knockout (KO) cell line using 293T cells is a powerful tool for scientific research. Here are some potential applications of this cell line in the fields of biotechnology and medical research: 1. Gene function studies: B2M (Beta-2-Microglobulin) is an important molecule in the immune system. By knocking out the B2M gene, its function in gene expression and cellular processes can be studied. Specific applications include confirming the role of B2M in disease models and understanding the mechanism of immune escape. 2. Drug screening and development: This cell line can be used to evaluate the effectiveness and toxicity of new drugs, especially in the process of drug development involving immune-related pathways. It can provide a background without a specific immune response for drug screening. 3. Cellular immunotherapy: It can be used to develop and test new cell immunotherapies, such as CAR-T cell therapy. Using this cell line, researchers can develop more targeted and effective treatments. 4. Tumor research: Since B2M plays an important role in certain types of cancer, by using B2M knockout 293T cells, we can gain a deeper understanding of the behavior of cancer cells and the potential points of action of treatment methods, which will help develop new cancer therapies. 5. Disease model construction: This cell line can be used to construct human disease models, especially diseases involving the immune system, such as autoimmune diseases. Researchers can use this model to explore the mechanism of disease occurrence and development, as well as screen potential therapeutic drugs.
Customer Q&As
What is the recommended growth medium? Does it require antibiotic selection?

A: DMEM supplemented with 10% fetal bovine serum.
It is not required to add the selection antibiotics when culturing the KO cells.

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
Helpful

β2M knockout cells are cells that have been genetically engineered to lack the expression of β2M protein. The cell line is very helpful for our research.

United States

08/15/2021

Invaluable tool

Great buy! By knocking out the β2M gene, we can gain a better understanding of how immune cells interact with each other and regulate immune responses.

United Kingdom

10/09/2021

Reliable service

The team is available to assist me with any questions about CRISPR knockout cell lines. I feel very good!

United States

02/23/2023

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