iLiteTM alphabeta - Instructions for Use (Abridged)

Important Note: Refer to Instructions for Use supplied with the kits when assay is being performed.

Receipt and Storage

Upon receipt, the kit should be stored at minus 80ºC .

Kit Contents

Each kit contains the following:

  • 1x Vial Cells
  • 1x vial IFN Stock
  • 1x vial IFN Positive Control and 1x vial IFN negative Control
  • 1x bottle Diluent
  • 1x bottle substrate and 1x bottle lysis buffer
  • 1x 96 well microwell plate for luminescence
  • 1x Instructions for Use

Handling/Disposal

This kit contains a stable transfected cell line of human origin. For the purposes of handling and disposal this material should be considered as a potentially hazardous biological material, Class 1 GMO and treated accordingly. Autoclave after use.

The assay has been calibrated based on the international reference standard for human interferon alpha (Hu-IFN-a 2b) provided by the UK National Institute for Biological Standards and Control (95/566).

Assay Procedure

To ensure kit performance the protocol should be reviewed in its entirety prior to use.
  1. Construct a standard curve from 0-100 IU/ml by preparing a series of dilutions using the IFN stock provided.
  2. Transfer 25ml of cell suspension to each well.
  3. Add 50ml of assay diluent to each well.
  4. Place 25ml of the IFN standards Negative Control, Positive Control or Unknown samples in individual wells.
  5. Incubate at 37oC in 5% CO2 for 4-18 hrs.
  6. After 18 hours add 100µl of the reconstituted Lysis substrate (refer to MSDS document) per well, and read on a luminometer, between 5 (min) and 30 (max) minutes after addition of lysis substrate.
  7. Construct a standard curve by plotting IFN activity on X -axis (log scale) and Relative Light Units (RLU) on Y scale (linear scale).
  8. Read Controls and Unknown samples from the standard curve or alternatively use a trendline equation to determine observed activity values.

The following standard curve for Human Type I IFN activity is provided as a demonstration only and should not be used to obtain test results.