The Protein and Enzymes Technical Committee of American Association of Cereal Chemists initiated a collaborative study to confirm whether the G12 antibody-based sandwich ELISA test kit is able to detect gluten in the lower mg/kg (ppm) level. Twenty laboratories investigated 24 heat-treated and non-heat-treated blind-coded samples with incurred gluten levels up to 100 mg/kg. The method has been validated for testing foods to conform to the defined Codex thresholds for gluten in gluten-free products at less than 20 mg gluten/kg. The collaborative study showed that low levels of gluten could be detected by G12 Sandwich ELISA with reproducibility RSDR of 32% and repeatability RSDr of 16%. Incurred samples showed a recovery between 62 and 135%. It is recommended that the method be accepted by AOAC as Official First Action.