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Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Calu-3 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate, respectively). 图2:磷酸化或非磷酸化的裂解中蛋白含量和450nm吸光值之间的关系。Calu-3细胞在85%满时收样,并用加入或未加磷酸酶抑制剂的裂解液进行处理(磷酸化或非磷酸化)。
Figure 1: Constitutive phosphorylation of DDR1 in Calu-3 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863 (upper, right). In contrast, a low level of phospho-DDR1 protein is detected in Calu-3 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total DDR1 protein from both nonphospho and phospho lysates are detected by PathScan® Total DDR1 Sandwich ELISA Kit #7845 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-DDR1 (Tyr792/796/797) rabbit antibody (right) or a total DDR1 rabbit antibody (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4. 图1:在磷酸酶抑制剂*存在时(磷酸化的裂解液),用PathScan® Phospho-DDR1 (panTyr) Sandwich ELISA Kit #7863能够检测到Calu-3细胞中磷酸化DDR1的组成型表达(上,右)。相比之下,没有磷酸酶抑制剂*存在时(非磷酸化的裂解液),仅能检测到低水平的磷酸化DDR1的表达。用PathScan® Total DDR1 Sandwich ELISA Kit #7845则能在磷酸化和非磷酸化的裂解液中检测到近似水平的总DDR1的表达(上,左)。上图显示450nm处的吸光值,下图显示用抗磷酸化DDR1 (Tyr792/796/797)的兔源单克隆抗体(右)或抗总DDR1的兔源单克隆抗体(左)进行免疫印迹检测的结果。*磷酸酶抑制剂中包含sodium pyrophosphate、β-glycerophosphate和 Na3VO4。