A colorimetric assay kit for the measurement of the total antioxidant status. Each kit is suitable for performing a minimum of 50 assays.

Note: Spectrophotometer equipped with a temperature-controlled cuvette holder is strongly recommended for use with this kit.

Substrate, Chromogen, Standard, Buffer, and a user protocol.

Toxicity: Standard Handling (A)

The Calbiochem Total Antioxidant Status Assay Kit is designed to assay antioxidant levels in serum or plasma samples. Additionally, the assay may be used to measure the antioxidant potential of (suitably solubilized) food and drug samples.

• Chromogen: Add 10 ml Buffer to each vial of Chromogen needed for the assay (10 tests per vial). After reconstitution, the Chromogen is stable for 2 days at 4°C or 8 h at room temperature.

• Substrate: Dilute each vial of Substrate needed for the assay by adding 7.5 ml Buffer. After dilution, the substrate is stable for 24 h at 4°C. NOTE: Some automated methods do not require substrate dilution.

• Standard: Add 1 ml distilled water to each vial of Standard needed for the assay. After reconstitution, the Standard is stable for 2 days at 4°C or 1 month at -20°C.

Upon arrival store the entire kit contents at 4°C. Note: The concentration of the standard is lot-specific. Please refer to the vial label for lot-specific concentration.

Calculation of Total Antioxidant StatusThe antioxidant concentration of the Standard is used to calculate the antioxidant levels in the samples.

1. Determine the ΔA for the samples, standard, and blank. ΔA = A - A_o
2. Calculate the concentration of antioxidants in the sample using the formula below:
<div class="Bio_doc_image">Figure 2: Antioxidant Concentration Formula
</div>
NOTE: If the antioxidant concentration is greater than 2.5 mM dilute the sample with 0.9% NaCl and re-assay. Typical antioxidant levels in normal human plasma are 1.30-1.77 mM.

Miller, N.J., et al. 1993. Clin. Science 84, 407.

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany