Transfer buffers may have become contaminated. Contamination can be transferred to the blots from electrophoresis and related equipment used in blot preparation. Post-antibody washes may not have been performed for a sufficient period of time or were not performed in a high enough volume. The blocki...
Wash the membrane in PBST for 10 min for three times and apply the diluted conjugated secondary antibody in blocking buffer (as per manufacturer’s instructions) and incubate 1 hour at room temperature. Wash the blot in PBST three times for 10 min each. Detection Apply the detection reagent ...
Western Blot即蛋白免疫印迹( Western Blot) 通过凝胶电泳将细胞或组织中的蛋白质转移到膜上,然后通过特异性抗体对电泳分离后的蛋白质进行着色,通过分析着色的位置和着色深度获得特定蛋白质在所分析的细胞或组织中表达情况。二、简单步骤: 1.蛋白的提取与定量。 2.跑胶电泳:一般用的是SDS-PAGE(十二烷基硫酸钠-聚丙...
产品名称:Western blot二抗稀释液说明书 规格:50ml 分类:免疫印迹 储存条件:4℃,6个月 用途:二抗稀释,可以防止抗体效价下降,减少抗体在组织上的非特异性吸附 注意事项:主要由TBS、BSA、防腐剂等组成。 特点: 1、采用全进口原料:高效、灵敏、特异,严格引进国际技术标准进行批量生产。
electrical current. The membrane can then be processed with primary antibodies specific for target proteins of interest. Next, secondary antibodies bound to enzymes are applied and finally a substrate that reacts with the secondary antibody-bound enzyme is added for detection of the antibody/protein ...
Note:If you are using a secondary antibody conjugated to HRP, do not use solutions containing NaN3from this point on. Incubate the secondary antibody in PBS with 1% BSA and 0.1% Tween-20 for 1 hour at room temperature with gentle agitation. ...
多克隆抗体（polyclonal antibody, pAb）：用一种包含多种抗原决定簇的抗原免疫动物，可刺激机体多个 B ...
Afterward, the membrane is washed and a secondary antibody is introduced that binds to the first. Oftentimes, a luminescent agent is used to assist scientists with the identification process. The material is washed again to remove any unbound items. Analysis of the size of the stained bands ...
(such as peroxidase) that is bound to the secondary antibody. This converts the soluble dye into an insoluble form of a different color that precipitates next to the enzyme and thereby stains the membrane. Development of the blot is then stopped by washing away the soluble dye. Protein ...
Signal detection is the next step within the western blot workflow. This step allows for the quantification of your protein of interest within your sample. The signal detection method depends on your selection of secondary antibody reporter. For example, if using an enzymatic reporter, at this poi...