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Buhlmann Assay pilot
Ran Buhlmann assay for first time using buffy coats from Tom Kadlik (RBC) and MGH. Used five dilutions each of Milk and Peanut allergen (1.00, 0.20, 0.040, 0.0080, 0.0016 µg/mL)
Followed Buhlmann protocol with following differences:
Heavy clumping in both the Kadlik and MGH samples, possibly due to sitting in staining buffer overnight. Kadlik samples were jettisoned due to low basophil event count. MGH 5/18/11 basophils were nonresponding to anti-FCeRI Ab - next time, should also include anti-IgE Ab as additional positive control. Samples were unsensitized to both milk and peanut series.