Cat.No. : CSC-RO0666 Host Cell: HEK293T
| Cat. No. | CSC-RO0666 |
| Description | This cell line is engineered to overexpress human leukocyte immunoglobulin like receptor B4 (LILRB4). |
| Gene | LILRB4 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293T |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Studying the interactions between immune cells and cancer cells 2. Studying the mechanisms of resistance to immune checkpoint blockade 3. High-throughput screening 4. Drug target validation |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| 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 |
Acute monocytic leukemia (AML) is a highly aggressive subtype of leukemia, characterized by the expression of immune checkpoint proteins like leukocyte immunoglobulin-like receptor B4 (LILRB4). This receptor plays a crucial role in immune evasion, making it a significant target for novel immunotherapies. In recent research, a humanized monoclonal IgG1 LILRB4-blocking antibody (h128-3) demonstrated promising results by improving immune regulation while reducing LILRB4 expression by 40–60%. However, its effectiveness was influenced by the interaction between the antibody's Fc region and Fc gamma receptors (FcγRs), highlighting the importance of this pathway in therapeutic design. The study further explored the mechanism of FcγR-dependent antigenic modulation, elucidating how inhibiting or removing FcγRI impacted the blocking function of h128-3, thus affecting T-cell-mediated cytotoxicity.
Figure 1. The researchers employed flow cytometry to analyze LILRB4 and FcγRI expression in HEK293T LILRB4/FcγRI overexpressing cells, assessing h128-3-induced LILRB4 internalization in monocytic AML. (Morse JW, et al., 2023)
Creative Biogene's Human LILRB4 Stable Cell Line - HEK293T offers a reliable platform for researchers investigating immune checkpoint mechanisms in myeloid malignancies. This cell line can be genetically modified using lentivirus to express human LILRB4, either alone or in conjunction with FcγRI. The cells are rigorously selected to ensure stable expression, providing a valuable tool for studying the functional dynamics of LILRB4 and FcγR interactions in vitro.
A: The Human LILRB4 Stable Cell Line - HEK293T provides a robust model for investigating the role of LILRB4, an immune inhibitory receptor, in immune checkpoint regulation. This cell line allows for the detailed analysis of LILRB4 signaling pathways, ligand interactions, and its impact on immune cell function, particularly in the context of cancer immunotherapy. The use of a stable HEK293T cell line expressing LILRB4 facilitates consistent and reproducible results, essential for advancing our understanding of immune checkpoint mechanisms and developing novel therapeutic interventions.
A: Researchers can employ the Human LILRB4 Stable Cell Line - HEK293T in high-throughput screening assays to identify antibodies or small molecules that bind to LILRB4, potentially modulating its activity. These screenings involve measuring changes in downstream signaling events, such as calcium flux, phosphorylation states of signaling molecules, or reporter gene activation, in response to compound treatment. Such studies are critical for discovering new drugs that can either enhance or inhibit LILRB4 signaling, offering therapeutic options for diseases where LILRB4 plays a key regulatory role.
A: To maintain the stability and expression of LILRB4 in the Human LILRB4 Stable Cell Line - HEK293T, it's crucial to culture the cells under optimal conditions, including the correct temperature (37°C), CO2 levels (5%), and using a medium supplemented with the appropriate selective antibiotics. Periodic verification of LILRB4 expression through flow cytometry or Western blot analysis is recommended to ensure consistent expression. Additionally, avoiding overgrowth and subculturing cells before reaching confluency will help maintain the cell line's integrity and expression levels.
A: The Human LILRB4 Stable Cell Line - HEK293T can significantly contribute to immune-oncology research by facilitating studies on how tumor cells exploit LILRB4 to evade immune detection and destruction. By expressing LILRB4 on HEK293T cells, researchers can mimic tumor cell interactions with immune cells, studying how LILRB4 engagement suppresses immune responses. Investigating the blockade or enhancement of LILRB4 signaling in this cell line could uncover new strategies for cancer immunotherapy, aiming to boost the immune system's ability to recognize and eliminate tumor cells.
A: To study ligand-receptor interactions involving LILRB4, researchers can employ several experimental approaches using the Human LILRB4 Stable Cell Line - HEK293T, including co-immunoprecipitation (Co-IP) to identify physical interactions between LILRB4 and its ligands. Surface plasmon resonance (SPR) and bio-layer interferometry (BLI) are additional biophysical methods that can quantify the binding affinity and kinetics of these interactions. Furthermore, functional assays, such as reporter gene assays or cytokine release assays, can assess the biological consequences of ligand binding to LILRB4, providing insights into how these interactions regulate immune responses.
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The Human LILRB4 Stable Cell Line - HEK293T ensures accurate and consistent expression of the LILRB4 receptor. This accuracy is critical for reliable investigation of LILRB4's role in immune modulation and receptor signaling, providing a solid basis for experimental reproducibility.
HEK293T cells, known for their high transfection efficiency, make the Human LILRB4 Stable Cell Line - HEK293T exceptionally receptive to further genetic modifications. This trait is beneficial for us looking to introduce additional genes or reporters into the cell line.
The Human LILRB4 Stable Cell Line - HEK293T benefits from the robust growth characteristics of HEK293T cells, facilitating easier scaling of experiments. This robust growth is advantageous for high-volume experimental setups, such as drug screening or large-scale studies.
Stable expression of LILRB4 in the Human LILRB4 Stable Cell Line - HEK293T enhances the reliability of data, especially in studies focusing on the immune checkpoint functions of LILRB4. Reliable data is crucial for forming valid conclusions and advancing scientific knowledge.
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