Pierce™ Silver Stain for Mass Spectrometry - FAQs

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10 product FAQs found

我使用用于质谱分析的Thermo Scientific银染染色剂染色后，得到的背景非常高。这是为什么？

以下是可能原因和解决方案：

- 染色后的凝胶过度显色： 缩短显色时间。
- 洗涤步骤缺漏或水质较差： 不要跳过和减少洗涤步骤，应使用超纯水洗涤。
- 所用设备被污染： 使用超纯水将设备冲洗干净。
- 制胶所用化学品不纯或预制胶已过期： 使用分析级化学品或使用未过期的预制胶。
- 终止液不能有效终止凝胶显色： 制备新的5%乙酸，并在凝胶孵育的最初几分钟里更换2次。

我使用用于质谱分析的Thermo Scientific银染染色剂染色后，条带很微弱或完全看不见。我该怎么办？

以下是可能原因和解决方案：

- 显色时间不足：凝胶显色时间>5分钟，或加入新配制的显色工作液。
- 样品中的蛋白质含量极低或无蛋白： 检测原始样品中的蛋白浓度。
- 溶液配制不恰当或遗漏了某些步骤： 检查溶液配制方法，并遵循操作步骤。
- 显色步骤前，水洗过度： 为完全去除之前使用的溶液，应洗涤凝胶3次，每次10分钟；从步骤2开始，再次染色。使用显色剂孵育前，不要过度洗涤。

进行质谱分析时，应如何处理经过质谱分析用Thermo Scientific银染染料染色的凝胶？

使用质谱分析用Thermo Scientific银染染料染色后，有多种方法可回收和处理蛋白质以用于质谱分析。对于胶内胰蛋白酶消化，无论是否进行还原和烷基化，均可使用Thermo Scientific胶内胰蛋白酶消化试剂盒（货号89871）。应考虑遵循质谱分析中核心设备的建议。

质谱分析用Thermo Scientific银染染料的灵敏度是多少？

质谱分析用Thermo Scientific银染染料具有次纳克级别的灵敏度，可在固定后30分钟内检测到蛋白含量低至0.25 ng的条带。

用于质谱分析的Thermo Scientific银染染料应如何保存？保质期是多久？

质谱分析用Thermo Scientific银染染料可在室温下稳定保存1年。

I used the Thermo Scientific Silver Stain for Mass Spectrometry and am getting very high background. Why is this?

Here are possible causes and solutions:

- Stained gel was overdeveloped. Reduce development time.
- Washing step(s) was missed or poor quality water was used. Do not skip or reduce wash steps and use ultrapure water.
- Contaminated equipment was used. Use clean equipment rinsed with ultrapure water.
- Impure chemical was used for gel preparation or precast gel has expired. Use analytical grade chemicals or use precast gels that have not expired.
- Stop solution was not effective in halting development of gel. Prepare new 5% acetic acid and replace it twice in the first minutes of incubation with the gel.

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

I used the Thermo Scientific Silver Stain for Mass Spectrometry and my bands are very faint or not visible at all. What should I do?

Here are possible causes and solutions:

- Insufficient development time. Develop gel for more than 5 minutes or add freshly prepared Developer Working Solution.
- Minimal or no protein present in sample. Check protein concentration in the original sample.
- Improper solution preparation or skipped steps. Check solution preparation and follow procedure.
- Excessive water wash before development step. Wash gel three times for 10 minutes each to completely remove the previous solutions and redo the staining procedure starting at step 2. Do not over wash prior to incubation in the developer.

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

How should I process gels stained with Thermo Scientific Silver Stain for Mass Spectrometry, for mass spectrometry analysis?

After using the Thermo Scientific Silver Stain for Mass Spectrometry, proteins can be recovered and processed by several methods for mass spec analysis. For in-gel trypsin digestion, with or without reduction and alkylation, use the Thermo Scientific In-Gel Tryptic Digest Kit (Cat No. 89871). Consider following the recommendations of the core facility that will be performing the mass spec analysis.

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

What is the sensitivity of the Thermo Scientific Silver Stain for Mass Spectrometry?

The Thermo Scientific Silver Stain for Mass Spectrometry provides subnanogram sensitivity, detecting proteins at less than 0.25 ng per band in 30 mins after fixing.

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

How should I store the Thermo Scientific Silver Stain for Mass Spectrometry and what is its shelf life?

We recommend storing the Thermo Scientific Silver Stain for Mass Spectrometry 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.