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Human CALR Knockout Cell Line-HEK293

Human CALR Knockout Cell Line-HEK293

Cat.No. :  CSC-RT2721

Host Cell:  HEK293 Target Gene:  CALR

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

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2721
Description This cell is a stable cell line with a homozygous knockout of human CALR using CRISPR/Cas9.
Target Gene CALR
Host Cell HEK293
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.
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.

Ship Dry ice
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Background

Applications

Publications

Q & A

Customer Reviews

The calreticulin (CALR) gene encodes a multifunctional protein called calreticulin, also known as calmodulin, CRP55, and endoplasmic reticulum-resident protein 60 (ERp60). This protein is primarily found in the endoplasmic reticulum (ER). As a soluble protein that binds calcium ions (Ca2+), calreticulin plays a crucial role in calcium homeostasis, binding Ca2+ with low affinity but high capacity. This property is essential for maintaining intracellular levels of calcium ions (second messengers in signal transduction), contributing to various signaling pathways. In addition, calreticulin is integral to the immune response, particularly in the production of major histocompatibility complex (MHC) class I proteins. During the synthesis of these proteins, calreticulin, along with other molecules such as ERp57 and tapasin, contributes to the assembly of MHC class I molecules, ensuring that they are properly folded and loaded with peptides to present antigens on the cell surface. This process is critical for the immune system's ability to recognize and fight pathogens. The presence and function of calreticulin is of great clinical significance. It is recognized by autoantibodies in autoimmune diseases such as systemic lupus erythematosus and Sjögren's syndrome, although it is not the major autoantigen in these diseases. More importantly, CALR gene mutations are found in a subset of myeloproliferative neoplasms (MPNs), particularly in patients without JAK2 or MPL gene mutations.
Applications of Human CALR Knockout Cell Line-HEK293 1. Cancer Research: The Human CALR Knockout Cell Line-HEK293 is extensively used in cancer research, particularly in studying the role of calreticulin (CALR) in various cancers. The absence of CALR allows researchers to understand its function in cell proliferation, apoptosis, and tumorigenicity. 2. Protein Folding Studies: CALR is a crucial protein involved in the proper folding of newly synthesized proteins in the endoplasmic reticulum. By utilizing the CALR knockout cell line, scientists can investigate the specific pathways and mechanisms affected by the lack of CALR, providing insights into diseases caused by protein misfolding, such as neurodegenerative disorders. 3. Immunology Research: CALR is known to play a role in the presentation of antigens to the immune system. The knockout model helps in examining the repercussions of its absence on immune cell activation, cytokine production, and immune evasion by pathogens or tumor cells. 4. Drug Development and Screening: In drug development, the Human CALR Knockout Cell Line-HEK293 serves as a critical tool for screening potential therapeutic agents targeting pathways involving calreticulin. This can aid in discovering new drugs for diseases related to CALR dysfunction, and also in evaluating the efficacy and mechanism of action of potential drugs. 5. Biogenesis and Degradation of Proteins: The absence of CALR in the Human CALR Knockout Cell Line-HEK293 enables researchers to explore its role in the biogenesis and degradation of glycoproteins.
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
Great!

Great! Utilizing the CALR knockout cell model can provide new avenues for understanding cellular processes and potential therapeutic targets for various diseases.

United States

10/09/2021

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