Invivogen
Menu

B16-Blue™ ISG-KO-STING Cells

B16-Blue™ ISG-KO-STING Cells Unit size Cat. code Docs Qty Price
Murine B16 melanoma - STING Knockout IRF-reporter cells
3-7 x 10e6 cells
bb-kostg
+-
$1,421.00

You may also need : QUANTI-Blue™ | View more associated products

STING Knockout IRF-Inducible SEAP Reporter B16 Melanocytes

B16-Blue™ ISG-KO-STING cells can be used for the study of the STING signaling pathway and for the detection of bioactive murine types I and II IFNs by monitoring the activation of the JAK/STAT pathway.

B16-Blue™ ISG-KO-STING cells were generated from the B16-Blue™ ISG cell line, a murine B16- F1 melanoma-derived cell line, through stable knockout of the STING gene. They express the secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of the I-ISG54 promoter which is comprised of the IFN-inducible ISG54 promoter enhanced by a multimeric ISRE.

B16-Blue™ ISG-KO-STING cells do not respond to cytosolic DNA, DMXAA, canonical and non-canonical CDNs while retaining the ability to respond to type I and type II IFNs.

Stimulation of these cells with IFN triggers the activation of the I-ISG54 promoter and the production of SEAP. Levels of SEAP in the supernatant can be easily determined using QUANTI- Blue™, a reagent that turns purple/blue in the presence of SEAP, by reading the OD at 620-655 nm.

B16-Blue™ ISG-KO-STING cells are resistant to Zeocin®.

Figures

Validation of IRF5 knockout by Western blot (Wes™)
Validation of IRF5 knockout by Western blot (Wes™)

Analysis of lysates from the B16-Blue™ (WT) and B16-Blue™ KO-STING (KO) cells using Anti-STING, followed by an HRP-conjugated anti-rabbit secondary antibody. The arrow indicates the expected band for the STING protein (43 KDa).


Response of B16-Blue ISG and B16-Blue ISG-KO-STING to CDNs and IFN-β:
Cells were stimulated with 30 µg/ml of the cyclic dinucleotides, and 103  U/ml of mIFN-β. Cells were not permeabilized.
After 24h incubation, the levels of IRF-induced SEAP were determined using QUANTI-Blue™.

Back to the top

Specifications

Murine IFN Detection range
• mIFN-α: 10e2 - 10e4 IU/ml
• mIFN-β: 10e2 - 10e4 IU/ml
• mIFN-γ: 0.1 ng - 1 µg/ml

Antibiotic resistance: Zeocin®

Quality control:

  • STING knockout is verified by functional assays and DNA sequencing to confirm frameshift mutation/deletion
  • These cells are guaranteed mycoplasma-free

Growth medium: DMEM, 10% (v/v) heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 µg/ml Normocin™, 100 U/ml penicillin, 100 µg/ml streptomycin

 

This product is covered by a Limited Use License (See Terms and Conditions).

Back to the top

Contents

  • 1 vial containing 3-7 x 106 cells
  • 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)

Dry ice shipping Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

Back to the top

FAQ Cell Lines

Visit our FAQ Any questions about our cell lines ? Visit our frequently asked questions page

Back to the top
Customer Service
& Technical Support
Shopping cart is empty