hientrung

Hi everybody, I am studying PCA Test (Passive cutaneous anaphylaxis) in ICR mice according to the following method: step1:anti DNP-IgE (mono clonal antibody produce in mice) intradermal injection. Each mouse(25-30g) 50µl (40 µl PBS saline+ 10 µlIgE). Skill: Mix IgE with saline well by vortexing (not by ultrasonic wave) step2: After 48hours. Each mouse is tail vein injected with 0.2ml of (200µg DNP-HAS + 3%Evant blue in PBSsaline).1h before challenge, mouse is oral administration treatment sample. Skill: do not fasting mouse (other experiments fasting is required) Evant blue and DNP-HSA (dinitro phenyl albumin) are dissolved well in saline by using ultrasonicator. step3: 30minutes after tail vein injection, mouse is sacrificed and removed dorsal skin and measure the pigmented area(1*1cm). Skill: get exactly area of each dorsal skin sites. When following above protocol, my results were not good. The pigment of the site which was intradermally injected by IgE is not darker than the site which was intradermally injected by PBS saline. Could you show me what the wrong is ??? the wrong protocol or my wrong techniques ?? And which step during this process that I have to note? I am sure that I did tail vein injection well, and intradermal injection may be OK (there is one big spot appears in skin surface). I am in a hurry. Please, help...help...