ADCC CTLA-4 Target Cell Line, Human Raji Cells (CAT#: AD-P005-C) Datasheet

Description
CTLA-4 (Cytotoxic T Cell Lymphocyte Antigen 4; also known as CD152) is a type I transmembrane protein, expressed on the cell surface of activated conventional T cells and on immunosuppressive regulatory T cells (Tregs) Constitutive expression. CTLA-4 is an inhibitory immune checkpoint that prevents over-stimulation of T cells and host damage. It has a competitive binding effect on stimulating CD28 ligand (CD80/CD86).

Raji-hCTLA4 cells are developed from Raji cell line (a cell line derived from human B lymphocytes). Raji cells have been successfully used as target cells for CAR-T toxicity assays and human effect studies, such as antibody-dependent cytotoxicity (ADCC), which can be derived from peripheral blood mononuclear cells, natural killer (NK) cells or T cells Jurkat reporter cell.
Features
Surface expression markers and IC in Raji-hCTLA4 cells.
Surface expression markers and IC in Raji-hCTLA4 cells
Stable overexpression of human CTLA-4 gene
Characterized by many markers expressed on the cell surface, including B cell receptor (BCR), CD19 and CD20
Constitutive expression of various immune checkpoints (IC), such as CD27, CD70, CD80, PD-L1 and 4-1BBL
The stability of 20 channels after thawing has been verified
Applications
Used as a target cell line for ADCC analysis of Jurkat NFAT-CD16 cells
Used as a target cell line for NK or CAR-T cell cytotoxicity assay
Used to develop other functional assays
Size
3-7 x 10e6 cells
Contents
Two kit formats are provided:
Complete, everything you need to get started
Core, used with customer-defined antibodies and target cells
Species Reactivity
Human
Cell Background
Raji cell line
Sample
Works with both adherent and suspension cells.
Validation
Flow cytometry verifies the overexpression of CTLA-4
Use anti-human CTLA-4 monoclonal antibody and Jurkat NFAT-CD16 cells as target cells for functional testing in ADCC analysis
Guaranteed Mycoplasma Free
Growth Medium
IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum (FBS; 30 min at 56 °C), 10 μg/ml Blasticidin, Pen-Strep (100 U/ml-100 µg/ml)
Test Medium
IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated FBS, Pen-Strep (100 U/ml-100 µg/ml)
Cell Purity
>95%
Cell Viability
>90%
Sterility Testing
Creative Biolabs provides sterility testing in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.
Identity Testing
Identity testing is required for newly established cell lines. Isoenzyme analysis is used to confirm the identity of the species of a cell line. Alternative methods for identity testing include DNA fingerprinting, STR analysis and karyology.
Virological Safety Testing
A broad range of viruses is susceptible to affecting human cell lines. We can provide in vivo/vitro virus saftey assays by utilizing various animal systems. These viruses include: adventitious viruses, bovine viruses, human and simian viruses, porcine viruses, retrovirus and rodent viruses.
Genetic Stability Testing
We perform cell genetic stability studies under ICH guidelines. We can provide guidance on the appropriate testing program upon your requirements.
Mycoplasma Testing
Guaranteed mycoplasma-free
Shipping
Dry ice
Storage
-80°C for short-term storage, liquid nitrogen for long-term storage
Warnings
Avoid multiple freeze/thaw cycles
Research Use Only
Our recombinant cell are for research use only, not for diagnostic or therapeutic use.
Quality Control
The expression of human CTLA-4 has been verified by flow cytometry.
Anti-hCTLA4-hIgG1 antibody and Jurkat-NFAT Lucia™ CD16 reporter cell line have been used to verify the induction of antibody-dependent cellular cytotoxicity (ADCC).
The stability of the 20 channels after thawing has been verified.
Raji-hCTLA4 cells are guaranteed to be mycoplasma-free.

Figure ADCC assay with Raji-hCTLA4 cells
Incubate Raji-hCTLA4 cells with gradient concentrations of Anti-hCTLA4 or Anti-β-galactosidase (β-gal) mAb for 1 hour. Then the Jurkat NFAT-CD16 effector cells were incubated with the target cells for 6 hours. The activation of NFAT reflecting the induced ADCC response was evaluated by measuring Lucia luciferase activity in the supernatant using Luc. The percentage of maximum response normalized to the IgG1 isotype is shown.

All products and services are for Research Use Only. Do Not use in humans.

ONLINE INQUIRY

Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.

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