PageBlue™ Protein Staining Solution - FAQs

这最有可能是因为脱色时间不足。我们建议将膜放在30%乙腈/20%乙醇溶液中另外脱色5分钟。

这最有可能是因为蛋白转印出现问题。请确认转膜液和转印条件是正确的。

以下是可能原因和解决方案：

- 样品中无蛋白质：使用一个已知含量的纯蛋白质作为对照
- 样品中的蛋白质含量不足：增加总蛋白上样量
- 未将SDS从凝胶上完全去除：开始染色前，更彻底地洗涤凝胶

这最有可能是因为SDS的干扰。我们建议增加染色前的凝胶洗涤次数和/或洗涤液体积。也可使用25%异丙醇/10%乙酸溶液或12%三氯乙酸将凝胶脱色5分钟。

由于小分子量蛋白质（< 10 kDa）在染色期间容易从凝胶中浸出，因此，在使用PageBlue蛋白质染色溶液对凝胶染色前，需要使用戊二醛固定。其他常见的蛋白质固定剂（如乙酸、异丙醇、乙醇、三氯乙酸等）在此不适用，因为会将蛋白质在染色期间从凝胶中洗脱。请参见使用手册第3页的固定步骤。

染色步骤中的第一个洗涤步骤是用于去除凝胶中的十二烷基硫酸钠（SDS），因为SDS可干扰染色。非变性凝胶不含SDS，因此，用于非变性PAGE时可省略该步骤。

使用25%异丙醇/10%乙酸溶液或12%三氯乙酸（TCA）固定凝胶蛋白质15分钟，可提高染色灵敏度。

该产品对蛋白质检测的动态范围为5–500 ng，灵敏度约比传统Coomassie R-250染料强10倍。

PageBlue蛋白质染色溶液最多可重复利用3次，检测灵敏度不会明显降低。

PageBlue蛋白质染色溶液含有Coomassie G-250，可用于对聚丙烯酰胺凝胶和PVDF膜上的蛋白质进行染色。其他成分拥有专利权。

PageBlue蛋白质染色溶液推荐室温保存，可在室温下稳定保存1年。

This is most likely due to insufficient destaining time. We recommend destaining the membrane in 30% acetonitrile/20% ethanol solution for an additional 5 mins.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

This is most likely due to a problem with the western transfer. Please confirm that the transfer buffer and transfer conditions are correct.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Here are possible causes and solutions:

- No protein was present in sample. Load a known amount of purified protein as a control.
- Insufficient amount of protein in sample. Load more total protein in gel.
- SDS not completely removed from gel. Wash gel more extensively before staining.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

This is most likely due to SDS interference. We recommend increasing the number of washes and/or wash volumes before staining. Destaining the gel for 5 minutes with 25% isopropanol/10% acetic acid solution or 12% trichloroacetic acid will also help.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Small proteins (less than 10 kDa) are susceptible to leaching from the gel during the staining procedure and require fixation with glutaraldehyde before staining the gel with PageBlue Protein Staining Solution. Other common protein fixatives (e.g., acetic acid, isopropanol, ethanol, TCA, etc.) are not suitable for this purpose, as proteins will be washed out of the gel during the staining procedure. Please see procedure for fixation, mentioned on Page 3 of the manual.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The first wash step in the staining procedure is designed to remove sodium dodecyl sulfate (SDS) from the gel, because SDS interferes with staining. Native gels do not contain SDS and, therefore, this step can be omitted from native PAGE applications.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Fixing gel proteins with 25% isopropanol/10% acetic acid solution or 12% trichloroacetic acid (TCA) for 15 minutes can increase staining sensitivity.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

It can detect proteins with a dynamic range of 5-500 ng, which is approximately 10 times more sensitive than traditional Coomassie R-250-based dyes.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

The PageBlue Protein Staining Solution can be reused up to three times without noticeable loss in detection sensitivity.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

PageBlue Protein Staining Solution contains Coomassie G-250 for protein staining on polyacrylamide gels and PVDF membranes. The rest of the composition is proprietary.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

We recommend storing the PageBlue Protein Staining Solution at room temperature where it is stable for a year.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.