ADCC hVISTA Target Cell Line, Human Raji Cells (CAT#: AD-P009-C) Datasheet

Description
VISTA contains the V domain Ig of T cell activation inhibitor (VISTA), also known as programmed death 1 homolog (PD-1H), which is a member of the CD28/B7 protein superfamily. Similar to other immune checkpoints (IC), VISTA was found to be elevated on immune cells infiltrating the tumor microenvironment (TME). It is mainly expressed on antigen presenting cells (APC) and directly inhibits the activation and proliferation of T cells. In addition, VISTA is expressed on many T cell subpopulations, so it can be regarded as a ligand and a receptor at the same time, thus transmitting external and internal inhibitory signals to T cells [2]. PSGL-1 has been identified as a VISTA receptor. Interestingly, its participation selectively inhibits T cells in acidic pH, such as in TME [3]. Some people think that VISTA and PD-1 inhibit T cell function in a synergistic manner. Therefore, a combination therapy for the two ICs is considered very promising.

Raji-hVISTA cells are developed from the human Raji cell line (a cell line derived from human B lymphocytes). Raji cells have been successfully used as target cells for human effect studies, such as antibody-dependent cytotoxicity (ADCC), which can be used with peripheral blood mononuclear cells, natural killer (NK) cells or T cell-derived Jurkat reporter cells.
Features
Surface expression markers and IC in Raji-hVISTA cells
Surface expression markers and IC in Raji-hVISTA cells
Stable overexpression of human VISTA 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
Applications
Used as a target cell line for ADCC analysis on Jurkat NFAT-CD16 cells
Used as target cell line NK or CAR-T cell for cytotoxicity determination
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 to verify PD-L1 overexpression
Used as a target cell for functional testing of anti-human PD-L1 monoclonal antibody and Jurkat NFAT-CD16 cells 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 hVISTA has been verified by flow cytometry.
The induction of antibody-dependent cellular cytotoxicity (ADCC) has been verified using anti-hVISTA-hIgG1 antibody and Jurkat-NFAT Lucia™ CD16 reporter cell line.
The stability of the 20 channels after thawing has been verified.
Raji-hVISTA cells are guaranteed to be free of mycoplasma.

Figure ADCC assay with Raji-hVISTA target cells
Incubate Raji-hVISTA cells with gradient concentrations of Anti-hVISTA 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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