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文献和实验cells with 0.5% Triton X-100 for 10’ at room temp. Wash cells with PBS +0.2% Tween20 Block cells with blocking buffer for 15’ at 37°C (I use o.2% Tween20 and 10% serum) Stain cells with antibody of interest diluted
Immunostaining with Anti-Digoxigenin Alkaline Phosphatase
Atlternate BCIP/NBT (Roche, #11697471001, 20 Tabs, makes 10 mL/Tab, $67) Blocking Reagent (Roche #11096176001, 50 g, $104, or 10x Block Buffer #11585762001, 1 set, $147) Alternate blocking reagents: Nonfat dry milk, mouse serum, goat serum. Tris
FACS Analysis Using Peripheral Blood Cells
. (Alternatively, Becton-Dickinson sells a product called "FACS lysis buffer" that is used after the staining protocol to lyse RBCs and fix the cells.) Cells should be washed 2-3x with FACS buffer (PBS supplemented with either 1% BSA or 5% FBS and containing 0.05
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