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文献和实验Absorption Control in Immunohistochemistry Using Phospho-Peptides Immobilized on Magnetic Beads
importance to employ a control to determine that the staining pattern is specific. One of the frequently used controls in IHC is a so-called absorption control: phospho-specific primary antibodies are first incubated with a phospho-peptide immunogen to block
the beads in culture medium at concentration 2.0%.This is beads stock. Stored in darkness at 4℃. ---Assay 1, Controls and samples: Intact control (No staining) 1 well -Negative control (azide treated) 1 well -Normal control
-maleimide 染料与膜蛋白冰上孵育2. preclear:加入 control IgG 和 protein A/G beads,孵育,离心,吸取上清3. IP:先后加入抗体和 protein A/G beads 孵育,离心,保留 beads4. RIPA 清洗 beads,DTT 处理5. 煮样,蛋白电泳,转印6. 采集荧光信号——激光扫描7. 封闭,一抗孵育,HRP 二抗孵育8. 采集化学发光信号——胶片显影 小编:如果那个时候有 iBright,就可以同时进行成像了。条带获取更容易,且对比数据更具代表
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