IFN-γ Reporter B16 Cells

STAT1-SEAP reporter mouse melanoma

ABOUT

Murine Type II IFN Reporter Cells

B16-Blue™ IFN-γ cells are designed to monitor mouse type II interferon IFN-γ-induced JAK/STAT1 stimulation or inhibition through SEAP activation. This colorimetric cytokine bioassay can be used to screen activatory or inhibitory molecules, such as engineered cytokines and neutralizing antibodies, respectively.

Mouse IFN-γ reporter cells respond specifically to mouse (m) IFN-γ (see figures). They do not respond to human (h)  IFN-γ or mIFN-α/β.

 

Key features

  • Readily assessable STAT1-inducible SEAP reporter activity
  • Convenient readout using QUANTI-Blue™ Solution
  • No response to human IFN-γ
  • No response to type I IFNs (e.g.  IFN-α/β)

Applications

  • Therapeutic development
  • Drug screening
  • Release assay

IFN-γ is a pleiotropic cytokine with anti-viral, anti-tumor, and immunomodulatory functions. 

More details

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

SPECIFICATIONS

Specifications

Target

IFN-γ

Target species

Mouse

Tested applications

Detection of mouse IFN-γ

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Mouse melanoma
Reporter gene
SEAP
Detection method
Colormetric
Detection range

0.1 ng - 1 µg/ml

Antibiotic resistance
Zeocin®
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    B16-Blue™ IFN-γ Cells
  • Cat code: 
    bb-ifng
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Zeocin® (100 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

Cell line description

B16-Blue™ IFN-γ cells were generated from the murine B16 melanoma cell line by the stable transfection with the secreted embryonic alkaline phosphatase (SEAP) reporter under the control of the ISG54 promoter. This promoter comprises IFN-stimulated response elements (ISRE) that are recognized by the ISGF3 complex. The binding of IFN-γ to its receptor triggers a signaling cascade leading to the activation of STAT1 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.
 

Interferon-gamma background

Interferon-gamma (IFN-γ) is the sole member of the type II IFN family. It is secreted from CD4+ Th1 cells and activated NK cells. It plays a role in activating lymphocytes to enhance anti-microbial and anti-tumor effects [1-]. In addition, it plays a role in regulating the proliferation, differentiation, and response of lymphocyte subsets.

IFN-γ exerts its action by first binding to a heterodimeric receptor consisting of two chains, IFNGR1 and IFNGR2, causing its dimerization and the activation of specific Janus family kinases (JAK1 and JAK2) [4, 5]. Two STAT1 molecules then associate with this ligand-activated receptor complex and are activated by phosphorylation. Activated STAT1 molecules form homodimers and are translocated to the nucleus, where they bind IFN-stimulated response elements (ISRE) in the promoter of IFN-inducible genes.

 

1. Ivashkiv L.B., 2018. IFNγ: signalling, epigenetics and roles in immunity, metabolism, disease and cancer immunotherapy. Nat Rev Immunol. 18(9):545-558.
2. Shtrichman R. & Samuel CE., 2001. The role of gamma interferon in antimicrobial immunity. Curr Opin Microbiol. 4(3):251-9.
3. Sato A. et al., 2006. Antitumor activity of IFN-lambda in murine tumor models. J Immunol. 176(12):7686-94.
4. Platanias L.C., 2005. Mechanisms of type-I- and type-II-interferon-mediated signalling. Nat Rev Immunol. 5(5):375-86.
5. Schroder K. et al., 2004. Interferon-gamma: an overview of signals, mechanisms, and functions. J Leukoc Biol. 75(2):163-89.

DOCUMENTS

Documents

B16-Blue™ IFN-γ Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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