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文献和实验antibody. For antiphosphoCREB (IgG, 0.7 mg/ml), I use 1:5,000 - 1:20,000 dilution in 4% BSA in TBST. This should be titrated for optimal results. I incubate with primary Ab 3 hours - overnight at room temp with shaking.10. Remove the antibody solution
-purified goat anti-mouse IgG (Jackson Labs; 1 mg/ml stock used at 1:300 dilution) Texas Red-conjugated affinity-purified donkey anti-rabbit (Jackson Labs;1 mg/ml stock used at 1:200 dilution)
(adjust pH to 7.8 with Binding buffer; red color) to the Protein A column.Mouse antibodies of the IgG1 subclass do not have a high affinity for protein A. Purification on protein A beads using standard conditions will yield approximately 1/10
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