Pierce™ Power Stainer Welcome Pack - FAQs

查看更多产品信息 Pierce™ Power Stainer Welcome Pack - FAQs (22833SPCL)

34 个常见问题解答

我在使用Thermo Scientific Power Stainer对凝胶进行染色时,发现聚丙烯酰胺背景中存在与蛋白质不相关的非特异性斑点或污点。你们能否帮我排除故障?

以下是可能原因和解决方案:

- 凝胶洗涤不充分。小型凝胶至少水洗1次,共5分钟;中型凝胶至少洗涤2次,每次5分钟。当同时对2块小型凝胶染色时,每块小型凝胶应水洗2 x 5 分钟。
- 染色/脱色时间不足。将染色/脱色时间延长30秒至1分钟。或者,延长凝胶在水中的脱色时间。
- 凝胶和凝胶垫之间有气泡。安装染色装置时,使用滚筒或移液管赶走凝胶与凝胶垫之间的所有气泡。

我在使用Thermo Scientific Power Stainer对凝胶进行染色时,凝胶过度脱色。这是为什么?

这可能是因为染色时间过长。我们建议将染色时间缩短30秒或1分钟。

我在使用Thermo Scientific Power Stainer时,得到的脱色结果不一致。我该怎么办?

以下是可能原因和解决方案:

- 凝胶洗涤不充分。小型凝胶至少水洗1次,共5分钟;中型凝胶至少洗涤2次,每次5分钟。当同时对2块小型凝胶染色时,每块小型凝胶应水洗2 x 5 分钟。
- 染色/脱色时间不足。将染色/脱色时间延长30秒至1分钟。或者,延长凝胶在水中的脱色时间。
- 凝胶和凝胶垫之间有气泡。安装染色装置时,使用滚筒或移液管赶走凝胶与凝胶垫之间的所有气泡。

经Thermo Scientific Power Stainer染色的凝胶,能否用于免疫印迹分析?

不能。蛋白质在染色期间被固定到了凝胶上,因此,不能得到最佳的免疫印迹结果。免疫印迹分析应使用未染色的凝胶。

经Thermo Scientific Power Stainer染色的蛋白质条带能否用于质谱?

可以。其质谱分析结果与传统Coomassie染色的蛋白质条带相似。

经Thermo Scientific Power Stainer染色的凝胶可保存多久?

经Thermo Scientific Power Stainer染色的凝胶可在塑料保护纸或水中保存长达4小时,无信号损失。

经Thermo Scientific Power Stainer染色的凝胶,能否使用商业化Coomassie染料再次染色?

可以。按照生产商提供的实验方案,只需将凝胶放置于Coomassie染料中染色30-60分钟并脱色即可。

自制或商业化Coomassie染料能否用于Thermo Scientific Power Stainer?

不能。Thermo Scientific Power Stainer需使用经优化的专用快速染色液和脱色液。

常规Coomassie染料能否用于Thermo Scientific Power Stainer?

不能。该系统对常规Coomassie染料无效。

Thermo Scientific快速染色液使用的是什么染料?其原理是什么?

使用的是Coomassie R-250染料。在有铂涂层的阳极和不锈钢阴极加以最优化的电压后,Coomassie R-250染料的负电荷使其能够通过凝胶并与蛋白质结合。未结合的染料将继续电泳并离开凝胶基质,从而使背景脱色。

能否将免疫印迹滤纸用于Thermo Scientific Power Stainer?

不能。Thermo Scientific Power Stainer、快速染色液和脱色液要求使用小型和中型凝胶垫(每张凝胶垫有8层)。凝胶垫可作为快速染色液和脱色液的储备层。标准免疫印迹滤纸会导致染色不均匀和背景不一致。

Thermo Scientific快速染色试剂盒中的凝胶垫能否重复利用?

不能。为获得最佳结果,应使用新鲜配制的染色和脱色垫。每次染色后,应丢弃用过的凝胶垫。

一个Thermo Scientific快速染色试剂盒能对多少块小型和中型凝胶进行染色?

Thermo Scientific快速染色试剂盒所含材料足够用于30块小型凝胶(小型试剂盒)或15块中型凝胶(中型试剂盒)的染色。

使用Thermo Scientific Power Stainer进行染色的凝胶尺寸和厚度应为多少?

尺寸为7 x 8.4 mm的小型凝胶和8 x 13.5 cm的中型凝胶分别适用于小型和中型凝胶垫。最常用的凝胶厚度为1 mm。若使用厚度大于或小于1 mm的凝胶,将需要进行时间优化。

Thermo Scientific Power Stainer可同时对多少块凝胶进行染色?

Thermo Scientific Power Stainer Cassette可同时对1-2块小型凝胶或1块中型凝胶进行染色。

Thermo Scientific Power Stainer的灵敏度(检测上限和检测下限)是多少?

检测区间与商业化蛋白质染料相近(约0.01至50 µg纯蛋白)。

Thermo Scientific Stainer包含哪些组件?其工作原理是什么?

Thermo Scientific Power Stainer(货号22833)由带有激活版Staining Software的Thermo Scientific Power Station(货号22838)以及Thermo Scientific Power Stain Cassette(货号22836)组成。该系统专为聚丙烯酰胺凝胶中蛋白质的快速Coomassie染色脱色而设计。传统Coomassie染色技术需要一小时至一夜的时间来完成染色和脱色,才能得到理想结果。与Thermo Scientific中型和小型凝胶快速染色试剂盒一起使用时,Thermo Scientific Power Stainer只需6分钟即可带来等同或优于传统Coomassie染色技术的染色效率。该系统将蛋白质固定在凝胶中,并通过电泳使带负电荷的Coomassie R250染料快速通过凝胶基质,从而实现蛋白质染色时间的显著缩短。染料通过聚丙烯酰胺并与蛋白质以离子键结合,产生亮蓝色条带和极低的背景。该系统已经过验证,对常用的预制和自制SDS-PAGE凝胶都适用。

I used the Thermo Scientific Power Stainer for staining of my gel. I see non-specific spots or splotches of stain in the polyacrylamide background not associated with protein. Can you help me troubleshoot?

Here are possible causes and solutions:

- Inefficient washing of gel. Wash Mini gel at least once for 5 mins and Midi gel twice for 5 mins each in water. When staining 2 Mini gels simultaneously, wash each Mini gel 2 x 5 mins in water.
- Insufficient staining / destaining time. Add additional 30 seconds to 1 minute of staining / destaining time. Or, destain the gel in water for additional time.
- Air bubbles trapped between gel and the pads. When assembling staining stack, use a roller or pipette to remove any air bubbles between the gel and the pads.

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

I used the Thermo Scientific Power Stainer for staining of my gel but my gel is over-destained. What happened?

This is likely due to the stain time being too long. We recommend decreasing the stain time by 30 sec or 1 min.

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

I am getting inconsistent destaining with the Thermo Scientific Power Stainer. What should I do?

Here are possible causes and solutions:

- Inefficient washing of gel. Wash Mini gel at least once for 5 mins and Midi gel twice for 5 mins each in water. When staining 2 Mini gels simultaneously, wash each Mini gel 2 x 5 mins in water.
- Insufficient staining/destaining time. Add additional 30 seconds to 1 minute of staining / destaining time. Or, destain the gel in water for additional time.
- Air bubbles trapped between gel and the pads. When assembling staining stack, use a roller or pipette to remove any air bubbles between the gel and the pads.

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

Can western blotting be performed on gels previously stained with the Thermo Scientific Power Stainer?

No. During the staining, the proteins are fixed to the gel thus the western blotting results would not be optimal. The unstained gel should be used for western blotting.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Are protein bands stained with the Thermo Scientific Protein Stainer compatible with mass spectrometry?

Yes. It is compatible with mass spectrometry with similar results to conventional Coomassie staining.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

How long can I store gels stained using the Thermo Scientific Power Stainer?

Gels stained using the Power Stainer can be stored in a plastic protective sheet or in water for up to 4 hours without loss of signal.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Can gels stained with the Thermo Scientific Power Stainer be re-stained with commercial Coomassie stain?

Yes. Just place the gel in the Coomassie stain for 30-60 minutes and destain according to the manufacturer's protocol.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Can homebrew or commercially available Coomassie stain be used with the Thermo Scientific Power Stainer?

No. The Thermo Scientific Power Stainer requires the optimized and proprietary Power Stain Solution and Destain Solution.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Can I use regular Coomassie stains with the Thermo Scientific Power Stainer?

No. The system will not work with regular Coomassie stain.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

What is the dye used in Thermo Scientific Power Stain Solution? How does it work?

Coomassie R-250 dye is used. When optimized voltage is applied to the platinum-coated anode and stainless steel cathode, the negative charge of Coomassie R-250 dye causes it to travel through the gel and bind to the proteins. Unbound dye continues through and out of the gel matrix to destain the background.

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

Can western blotting filter paper be used with the Thermo Scientific Power Stainer?

No. The Mini and Midi Gel Pads (8 layers per pad) are required to be used with the Thermo Scientific Power Stainer, Power Stain Solution and Destain Solution. The gel pads act as reservoirs for Power Stain Solution and Destain Solution. Standard western blotting filter paper will cause uneven staining and patchy background.

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

Are the gel pads in the Thermo Scientific Power Staining Kits reusable?

No. For the best results, use freshly prepared staining and destaining pads. Discard the used pads after each staining.

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

How many Mini and Midi gels can be stained with each Thermo Scientific Power Staining Kit?

Thermo Scientific Power Staining Kits have enough material for 30 Mini gels or 15 Midi gels using the corresponding kit.

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

What gel sizes and thickness can be stained with the Thermo Scientific Power Stainer?

Mini gels with dimensions of 7 x 8.4 mm and Midi gels with dimensions of 8 x 13.5 cm are compatible with Mini and Midi gel pads. Gel thickness of 1 mm is the most common. Gels with a thickness greater or less than 1 mm would require time optimization.

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

How many gels can be stained simultaneously with the Thermo Scientific Power Stainer?

The Thermo Scientific Power Stainer Cassette accommodates the staining for 1 or 2 Mini gels or 1 Midi gel.

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

What is the sensitivity (upper and lower detection limits) of the Thermo Scientific Power Stainer?

Detection limits are similar to those of commercially available protein stains (approximately 0.01 to 50 µg of pure protein).

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

What are the components of the Thermo Scientific Power Stainer and how does it work?

The Thermo Scientific Power Stainer (Cat. No. 22833) consists of a Thermo Scientific Power Station (Cat. No. 22838) with activated Staining Software and a Thermo Scientific Power Stain Cassette (Cat. No. 22836). It is designed for rapid Coomassie staining of proteins in polyacrylamide gels and subsequent removal of stain from the gel background. Traditional Coomassie staining techniques require one hour to overnight of staining and destaining to achieve the desired results. When used in conjunction with Thermo Scientific Midi and Mini Gel Power Staining Kits, the Thermo Scientific Power Stainer is designed to provide staining efficiency in as few as 6 minutes that is equivalent to, or better than, traditional Coomassie staining techniques. This significant reduction in protein staining time is accomplished by fixing the protein to the gel and electrophoretically transporting the negatively charge Coomassie R250 dye rapidly through the gel matrix. The dye passes through the polyacrylamide and ionically binds to the protein, resulting in crisp blue bands with minimal background. The system has been verified to work with commonly used pre-cast and homemade SDS-PAGE gels.

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