Shreffler:Notebook/Alex Daily/2011/05/26

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Buhlmann Assay pilot 3

Ran Buhlmann assay using Wayne's peripheral blood, drawn 5/26/11. 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:

  • Incubation step in incubator rather than water bath
  • Included anti-IgE Ab positive control


Observations/notes

Procedure went smoothly. Due to use of whole blood rather than buffy coat, pellet was quite small. Nonetheless, was able to collect 700 basophil events (2:30 flow time per sample). Wayne's basos were nonresponding to anti-FCeRI/IgE Ab, and were unsensitized to milk and peanut.