Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line

Name: Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line
SKU: CSC-RO0671
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-RO0671 Size : >1x10⁶ frozen cells/vial

Inquire for Price

Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No.	CSC-RO0671
Description	This cell line is engineered to stably co-overexpress human PDCD1 and CD81-GFP in a human CD274 knockout cell line.
Target Gene	PDCD1/CD81-GFP
Host Cell Species	Homo sapiens (Human)
Applications	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
Size	>1x10⁶ frozen cells/vial
Stability	Validated for at least 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Storage	Liquid nitrogen
Shipping	Dry ice

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

Quick Inquiry

Background

Applications

Q & A

Customer Reviews

The Human CD274 gene, also known as PD-L1, encodes a protein that is involved in the regulation of the immune response by interacting with the PD-1 receptor on T cells. PD-L1 is expressed on various cell types, including immune cells and tumor cells, and its interaction with PD-1 can lead to the inhibition of T cell activation, making it a critical player in immune evasion by cancer cells. Additionally, the PDCD1 (PD-1) gene encodes the PD-1 receptor, and CD81 is a component of the tetraspanin family that modulates cell signaling.

The stable cell line expressing PD-1 and CD81 with a GFP (Green Fluorescent Protein) tag allows for the visualization and study of these proteins in living cells. This cell line is instrumental for investigating the PD-1/PD-L1 signaling pathway, which is a key target for cancer immunotherapy. It enables researchers to study the dynamics of PD-1 and PD-L1 interactions and their effects on immune cell function, providing insights into the mechanisms of immune checkpoint inhibition.

The Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line is a unique cell line in which the expression of CD274 (PD-L1), PDCD1 (PD-1), and CD81 has been genetically modified to include GFP tags. This cell line is particularly useful for studying the interactions between immune cells and tumor cells, as well as the development of immunotherapies targeting the PD-1/PD-L1 pathway.

（1） Immune Evasion Mechanisms in Cancer:

PD-L1 is often overexpressed in cancer cells, allowing them to evade the immune system by binding to PD-1 on T cells. This cell line can be used to study the interactions between PD-L1 and PD-1, and to test the efficacy of drugs that block this pathway, such as immune checkpoint inhibitors.

（2） Development of Immunotherapies:

The CD274(-)/PDCD1/CD81-GFP Stable Cell Line is an excellent tool for screening compounds that can modulate the PD-1/PD-L1 interaction, aiding in the development of new immunotherapies for cancer treatment.

（3）Study of T Cell Activation and Inhibition:

By observing the GFP-tagged PD-1 and PD-L1 interactions, researchers can gain insights into the mechanisms of T cell activation and inhibition. This cell line can be used to study how the PD-1/PD-L1 pathway affects T cell function and to identify potential targets for modulating immune responses.

Customer Q&As

What methodologies are recommended for quantitatively analyzing the GFP signal intensity in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line to assess the co-expression levels of these markers?

A: For quantitative analysis of GFP signal intensity in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line, flow cytometry is highly recommended. This technique allows for the precise measurement of GFP fluorescence, thereby assessing the co-expression levels of CD274(-), PDCD1, and CD81 markers. Before proceeding, cells should be properly prepared and resuspended in a suitable buffer. Employing a flow cytometer calibrated for GFP detection ensures accurate measurement. Additionally, data analysis software can be used to analyze fluorescence intensity, providing insights into the expression levels of these proteins. To complement flow cytometry, fluorescence microscopy can be employed to visualize the distribution and localization of GFP-tagged proteins within the cells.

Which strategies should be implemented to enhance the transfection efficiency of plasmids in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line, particularly when studying gene overexpression or knockdown effects?

A: Enhancing transfection efficiency in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line, especially for studies focusing on gene overexpression or knockdown, requires optimization of several key parameters. Electroporation or lipid-based transfection methods should be carefully selected based on their compatibility with this cell line. Optimization involves adjusting the DNA or siRNA concentration, the volume of transfection reagent, and the ratio of DNA to reagent. Additionally, performing a pilot study to determine the optimal conditions for transfection without affecting cell viability is crucial. Post-transfection, the efficiency can be evaluated using quantitative PCR or Western blot for the target gene, ensuring that the modifications do not interfere with the baseline GFP expression or the cell line's stability.

What are the critical factors to consider when designing experiments using the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line for drug screening assays aimed at identifying inhibitors of the PD-1/PD-L1 pathway?

A: When designing drug screening assays with the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line to identify inhibitors of the PD-1/PD-L1 pathway, it's essential to consider several critical factors. First, the choice of assay format (e.g., high-throughput screening compatible) and detection method (e.g., fluorescence-based readouts) should align with the GFP expression. It's vital to establish baseline GFP fluorescence intensity to accurately measure any modulation upon drug treatment. Furthermore, including a range of drug concentrations to determine the dose-response curve is crucial for assessing drug efficacy. Control treatments, such as known PD-1/PD-L1 pathway inhibitors, should be included to validate the assay's sensitivity and specificity. Lastly, evaluating cell viability alongside the primary assay endpoints is necessary to distinguish between specific pathway inhibition and general cytotoxic effects.

How can the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line be leveraged to study the molecular interactions between PD-1 and its ligands, and what experimental setups are most effective for such analyses?

A: To study the molecular interactions between PD-1 and its ligands using the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line, co-immunoprecipitation (Co-IP) followed by Western blot analysis is highly effective. This approach allows for the detection of physical interactions between PD-1 and its ligands by immunoprecipitating PD-1 or its ligands from the cell lysates and probing with specific antibodies against the interaction partners. Additionally, fluorescence resonance energy transfer (FRET) can be utilized to study the proximity between PD-1 and its ligands in live cells, with GFP serving as a fluorescent reporter. These experiments can be complemented with functional assays, such as T-cell activation or inhibition assays, to understand the biological significance of these interactions.

What specific steps should be taken to ensure the maintenance of GFP fluorescence and the stability of transgene expression in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line during long-term culture?

A: Ensuring the maintenance of GFP fluorescence and stability of transgene expression in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line during long-term culture requires meticulous cell culture practices. This includes using a consistent culture medium and conditions recommended by the cell line provider, regularly monitoring GFP expression levels through flow cytometry or fluorescence microscopy, and avoiding over-confluency by subculturing cells at appropriate intervals. It's also advisable to freeze down multiple aliquots of cells at early passages to preserve the original expression characteristics. Periodic validation of transgene expression through molecular techniques such as qPCR or Western blotting for GFP and the target transgenes is recommended to confirm stability over time.

Ask a Question

If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.

Question *

Name

Country *

Email *

Customer Reviews

Clear Visualization of CD81

This cell line uniquely incorporates GFP (green fluorescent protein), which is fused to the CD81 protein, allowing for clear visualization under fluorescent microscopy. The Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line enables us to observe CD81 localization and trafficking in real time, providing insights into cellular dynamics and interactions.

United States

2023-03-14

Dual Protein Knockout

The Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line has both CD274 and PDCD1 knocked out, which allows for the study of immune checkpoint pathways without the interference of these proteins. This dual knockout feature is invaluable for us focusing on the independent roles of other immunological markers like CD81 in cellular processes.

United States

2022-02-05

Enhanced Experimental Control

By lacking CD274 and PDCD1, this cell line serves as an excellent control for studying the specific effects of therapies targeting these pathways. The Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line provides a clear baseline to assess the impact of manipulating other molecular targets in the context of immune modulation.

Canada

2023-07-16

Facilitates Complex Immune Studies

The specific configuration of this cell line—combining knockouts with a fluorescent marker—enables complex studies of immune interactions and functions. We can use the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line to dissect the multifaceted roles of CD81 in the immune system, particularly in the presence or absence of typical checkpoint inhibitions.

Germany

2021-11-12

Write a Review

Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.

Product Name *

Catalog Name *

Rating

Needs improvement

Satisfaction

General satisfaction

Very satisfaction

Product Review Title *

Summary *

Name

Country *

Email *

Related Products

Cat.No.	Product Name	Price
CSC-RO0062	Mouse TLR11 Stable Cell Line-HEK293T	Inquiry
CSC-RO0009	Human LAG3 Stable Cell Line-CHO-K1	Inquiry
CSC-RO0100	Human CTLA4 Stable Cell Line-HEK293T	Inquiry
CSC-RO0065	Mouse 4-1BB Stable Cell Line-CHO-K1	Inquiry
CSC-RO0080	Human CD28 Stable Cell Line-HEK293T	Inquiry
CSC-RO0013	Human HAVCR2 Stable Cell Line-CHO-K1	Inquiry
CSC-RO0075	Human VTCN1 Stable Cell Line-HEK293T	Inquiry
CSC-RO0112	Human CD274 Stable Cell Line-HEK293T	Inquiry
CSC-RO0012	Human CD274 Stable Cell Line-HEK293T	Inquiry
CSC-RO0046	Human TNFRSF4 Stable Cell Line-CHO-K1	Inquiry

Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line

Cell Line Information

Cell Culture Information

Safety and Packaging

Background

Applications

Q & A

Customer Reviews

What methodologies are recommended for quantitatively analyzing the GFP signal intensity in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line to assess the co-expression levels of these markers?

Which strategies should be implemented to enhance the transfection efficiency of plasmids in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line, particularly when studying gene overexpression or knockdown effects?

What are the critical factors to consider when designing experiments using the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line for drug screening assays aimed at identifying inhibitors of the PD-1/PD-L1 pathway?

How can the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line be leveraged to study the molecular interactions between PD-1 and its ligands, and what experimental setups are most effective for such analyses?

What specific steps should be taken to ensure the maintenance of GFP fluorescence and the stability of transgene expression in the Human CD274(-)/PDCD1/CD81-GFP Stable Cell Line during long-term culture?

Clear Visualization of CD81

Dual Protein Knockout

Enhanced Experimental Control

Facilitates Complex Immune Studies

Related Products

Related Services