Cat.No. : CSC-SC002309-1 Host Cell: HEK293
| Cat. No. | CSC-SC002309-1 |
| Description | This cell line is engineered to stably overexpress human CALR (calreticulin). |
| Gene | CALR |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| 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. |
| 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. |
| 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 |
Within exon 9 of the calreticulin (CALR) gene, 20% to 30% of instances of primary myelofibrosis (PMF) and essential thrombocythemia (ET) are caused by frameshifting (−1/+2) mutations. The researchers show that myeloproliferative neoplasms (MPNs) are driven by mutant CALR proteins, which are the product of a −1/+2 frameshifting mutation of CALR exon 9. These proteins interact with primary megakaryocytic progenitors as well as other cells to activate the thrombopoietin receptor (TpoR/MPL). Up to 160 ng/mL of mutant CALR is produced by patients and is present in their plasma. It frequently binds to soluble transferrin receptor 1 (sTFR1) to increase its stability. The recombinant mutant CALR promotes thrombopoietin-independent colony formation in cell lines via activating TpoR. Assays for bioluminescence resonance energy transfer verify that secreted mutant CALR specifically interacts with TpoR on target cells, causing the cells to become hypersensitive to exogenous mutant CALR.
Figure 1. The researchers plated HEK293T cells on ibidi μ-slides and transfected them with TFR1-mCherry and CALR-linker-GFP fusion proteins. The researchers demonstrated that sTFR1 acts as a carrier protein for mutant CALR, studying its interaction through immunoprecipitation (IP) with biotinylated antimutant CALR antibody and subsequent analysis via nontargeted MS. They also examined its stability with rhCALR-del52 in varying media, using ELISA and confocal microscopy to observe colocalization with TFR1 in HEK293T cells. (Pecquet C, et al., 2022)
A: The Human CALR Stable Cell Line - HEK293, due to its stable expression of CALR (Calreticulin), holds unique advantages in anti-tumor drug screening. CALR's expression in various tumor cells is closely related to tumor immune escape and inhibition of cell apoptosis. Therefore, this cell line can be used to evaluate whether anti-tumor drugs can enhance the immune visibility of tumor cells or induce tumor cell death by regulating CALR expression or function.
A: In the Human CALR Stable Cell Line - HEK293, overexpression of CALR may affect cellular calcium homeostasis and endoplasmic reticulum stress response, thereby impacting cellular metabolism. As a calcium-binding protein, overexpression of CALR could enhance the cell's capacity for calcium storage and release, alter intracellular signaling, and affect pathways related to cell growth, differentiation, and survival.
A: The Human CALR Stable Cell Line - HEK293, with its stable expression of CALR, can be used to study the mechanisms of immune checkpoint inhibitors, such as PD-1/PD-L1 inhibitors, in cancer treatment, especially in regulating the tumor immune microenvironment. The expression of CALR may affect the interaction between tumor cells and immune cells, thereby influencing the efficacy of immune checkpoint inhibitors.
A: Utilizing the CALR expression characteristic of the Human CALR Stable Cell Line - HEK293, one can study the presentation efficiency of CALR-related antigens in tumor vaccines and their impact on immune responses. By understanding the role of CALR in tumor immune escape, vaccines can be designed to more effectively activate the immune system's recognition and attack against tumors.
A: As the Human CALR Stable Cell Line - HEK293 stably expresses CALR, it serves as an ideal tool for studying the role of CALR in the apoptosis process. The expression of CALR is related to various apoptosis pathways, including endoplasmic reticulum stress-induced apoptosis and immune-mediated cell death. Therefore, this cell line can be utilized to delve into how CALR regulates these apoptosis pathways.
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The Human CALR Stable Cell Line - HEK293 achieves precise and efficient expression of CALR protein by stably integrating the CALR gene into HEK293 cells, offering an ideal tool for studying the function and mechanism of CALR.
Using the Human CALR Stable Cell Line - HEK293 allows for direct studies on CALR-related cellular functions and signaling pathways, speeding up the process of scientific discoveries.
Developed based on the human cell line HEK293, the Human CALR Stable Cell Line - HEK293 ensures high biocompatibility and relevance of experimental data to humans.
The Human CALR Stable Cell Line - HEK293 is suitable for a wide range of research fields, including immunology, cell death, and cancer research, offering broad application prospects.
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