Answers:

• Posted by Manoj Sharma on Wednesday, January 6, 2016

  Hi Lauren,

  We have revised the preparation as follows:

  B. Preparation of test samples
  1. One volume of cells or tissues can be homogenized in 4 volumes of the assay buffer (8448a). Centrifuge the samples at 13,000 ×g for 10 minutes to remove insoluble material.
  2. Prepare a fresh working buffer by mixing assay buffer (8448a), reductant (8448e) and sample reagents (8448d) at 93:2:5 ratio (volume ratio). (e.g., 930 μL mixing assay buffer, 20 μL reductant, and 50 μL sample reagents)
  3. Mix sample (cell lysate or serum) with equal volume of working buffer.
  4. Centrifuge the mixed sample 13,000 ×g for 5 minutes and take the supernatant as the test samples for the iron assay.
  5. Test samples should be serial diluted to make sure the readings are within the standard curve range. Prepare test samples to a final volume of 50 μL/well on the 96-well flat bottom plate.