iLiteTNF-alpha Assay Ready Cells

$980.00

The Eagle Bio iLite Assay Ready Cells are designed for the specific detection of drug potency in serum/plasma as well as neutralizing antibodies in serum/plasma. They utilize an assay technique called reporter gene assay ( RPG ) to analyze serum/plasma samples. The reporter genes utilized are encoded with a bioluminescent luciferase (Firefly luciferase). Different levels of luminescence detected from each reporter gene cell indicates different levels of expression. The iLite® TNF-alpha Assay Ready Cells are for research use only and not for use in diagnostic procedures.

iLite TNF-alpha Assay Ready Cells

For Research Use Only

The Eagle Biosciences iLite® TNF-alpha Assay Ready Cells are intended for :

      • Testing anti-TNF-alpha activity
      • Testing NAb anti-TNF-alpha activity

      Content: 1.5 ml of Assay Ready Cells suspended in RPMI 1640 with 20% heat inactivated fetal bovine serum (FBS), 10% glycerol, and 2.5% dimethyl sulfoxide (DMSO) .

      Storage: -80°C, Cells should be used within 30 min of thawing.


      Key benefits of iLite® Assays

      • Highly specific reporter gene cell lines
      • Very sensitive cell line responses (>10 fold inductions)
      • Assay Ready Cells – ready-to-use from the freezer, without culturing of cells
      • Assays within a workday (typically 4-7 hour assays)
      • Normalization gene, which eliminates unwanted matrix effects

Product Developed and Manufactured by Svar Life Science

Additional Information

Background

TNF-alpha promotes inflammatory responses, which, in turn, contribute to the clinical problems associated with many inflammatory disorders, including rheumatoid arthritis, ankylosing spondylitis, Crohn’s disease, psoriasis and refractory asthma. These diseases are sometimes treated with monoclonal anti-TNF-alpha antibody constructs, including infliximab (Remicade), adalimumab (Humira), etanercept (Enbrel), certolizumab pegol (Cimzia), or golimumab (Simponi). Prolonged therapies with these TNF-alpha inhibitors may lead to development of neutralizing antibodies (NAbs) which may counteract the TNF-alpha antagonist activity of the drugs. The iLite® TNF-alpha reporter cells are engineered cells for use in a normalized reporter gene bioassay for screening of anti-TNF-alpha activity in samples. The assay is a simple and easy test format which detects anti-TNF-alpha activity and, indirectly, the presence of anti-drug antibodies.

Assay Principle

The iLite™ TNF-alpha Assay Ready Cells are engineered cells optimized to express Firefly luciferase under the control of a TNF-alpha responsive promoter and Renilla luciferase under the control of a constitutive promoter. TNF-alpha binds to the TNF receptor on the cell surface and activates the TNF-alpha regulated Firefly luciferase reporter gene construct. The Firefly luciferase signal can be measured in a luminometer following addition and incubation of the luciferase substrate. The Firefly luciferase signal is proportional to the functional activity of TNF-alpha in the sample. In the presence of TNF-alpha inhibitor activity in test samples, the amount of free TNF-alpha is reduced, resulting in a decreased stimulation of Firefly luciferase production. The Firefly luciferase signal is inversely proportional to the amount of anti-TNF-alpha activity in the samples. The constitutively expressed gene Renilla luciferase can be sequentially measured in the same wells as the Firefly luciferase, following addition and incubation of the Renilla substrate. This allows for normalization of results by compensating for cell number variation and eliminating potential matrix effects from a serum sample, for example. In samples with no suspected matrix effect, the Renilla reading may be excluded.

Assay Quick Guide

Click here for the Assay Summary: Anti-TNF-alpha Drug Activity Protocol

Why Choose the iLite® TNF-alpha Assay Ready Cells?

  • iLite technology employs the cells’ inherent signaling pathways – a true model of cellular events
  • Assay Ready Cells – no culturing required!
  • No protein tagging or labeling required
  • Short assay times
  • TNF-alpha inhibitor activity: 30min + 3h
  • Neutralizing antibodies: 30min + 30 min + 3h
  • Highly reproducible between batches
  • Normalization gene allows for consistent results
  • Adjusts for serum matrix effects and differences in cell number

Assay Range

  • anti TNF Drug Assays: IFX Drug 10-40ng/mL, ALD Drug 5-30 ng/mL, ETN Drug 2-10 ng/mL
  • Neutralizing antibodies to anti TNF drug: 20 – 2560 Antibody Titer. Samples can be further diluted to detect higher titer samples, there is no maximum range of detection.

Frequently Asked Questions


Manual

Product Manual


Publications

References (using these cells)