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蛋白凝膠電泳及相關(guān)的Western-Blot是蛋白分析的主流技術(shù) (Figure 1)。Applied BioProbes 公司提供兩種蛋白染料： 1)Coomassie® Blue Fast Stain, 一種即用型的蛋白染料，無需甲醇或乙酸固定、或脫色； 2) eLuminol™ Protein Gel Stain，一種具有*靈敏度的蛋白熒光染料，用于檢測(cè)SDS-PAGE凝膠電泳中的總蛋白。Applied BioProbes 公司也提供一種專門檢測(cè)磷酸化蛋白的熒光染料： Phos-Tag™ Phosphoprotein Gel Stain。Applied BioProbes提供的 ECL Western Blotting Detection Kit用于WB檢測(cè)HRP標(biāo)記的抗體及其關(guān)聯(lián)的抗原，具有快速檢測(cè)、靈敏度*等特點(diǎn)。另外，Applied BioProbes也提供了兩種蛋白定量分析試劑盒： 1) Bradford Protein Assay Kit；和 2) BCA Protein Assay Kit。
Figure 1. Protein gel analysis workflow
蛋白染料選購指南
| Coomassie® Blue Fast Stain | eLuminol™ Protein Gel Stain | Phos-Tag™ Phosphoprotein Gel Stain | |
| 靈敏度 | 20 ng | 0.5 ng | 1 ng |
| 操作性 | Ready to use | Supplied as stock solution | Ready to use |
| 耗時(shí) | 2 hr (standard) 20 min (microwave) | 90 min (standard) 30 min (microwave) | 4 hr (standard) 2 hr (microwave) |
| 檢測(cè)方法 | 比色法 | 熒光 | 熒光 |
| 質(zhì)譜兼容 | 是 | 是 | 是 |
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Figure 2. Gel stain results with Coomassie® Blue Fast Stain (left), eLuminol™ Protein Gel Stain (middle), and Phos-Tag™ Phosphoprotein Gel Stain, followed by eLuminol™ total protein stain (right). Two-fold serial dilutions (from left to right) of protein marker were separated via SDS-PAGE and then stained with above stains.
| ECL化學(xué)發(fā)光檢測(cè)試劑盒 |
Figure 3. Detection of protein in Western Blots with ABP kit (left) and Millipore Kit (right).
| Bradford Protein Assay Kit | BCA Protein Assay Kit |
Figure 4. BSA standard curve with Bradford Protein Assay Kit (left) and BCA Protein Assay Kit (right).