SYPRO™ Red Protein Gel Stain (5000X in DMSO), 500 μL - FAQs

View additional product information for SYPRO™ Protein Gel Stains - FAQs (S12000, S12001, S6650, S6651, S12010, S12000X3, S6654, S6653, S21900)

6 product FAQs found

如果我将SYPRO Orange或SYPRO Red染色液的终浓度增加至1X以上，会增强染色蛋白质的信号吗？

不会。染料浓度高于1X不会带来更好的检测结果。相反，随着染料浓度的增加，背景荧光会增强，染料会发生自淬灭，从而降低信号。

经SYPRO Ruby、SYPRO Orange或SYPRO Red染色的凝胶能否干燥？

经过SYPRO染料染色的凝胶可放置在玻璃纸之间进行干燥，但有时会轻微降低灵敏度。如果在纸上干燥凝胶，光会散射且灵敏度会降低。不推荐使用其他塑料，因为通常使用的塑料是不可透过紫外光的。

我能否使用SYPRO Ruby、SYPRO Orange或SYPRO Red蛋白质凝胶染料对蛋白质进行预染，然后将蛋白质在凝胶上进行电泳？

不能。上样溶液含有很多SDS，因此，SYPRO Ruby、SYPRO Orange和SYPRO Red染料会只与游离的SDS结合，而极少与蛋白质结合。在电泳前，可预先使用ATTO-TAG CBQCA（货号A2333）、DDAO琥珀酰亚胺酯（货号C34553）或TAMRA-琥珀酰亚胺酯（货号C2211）染料或TC-FLAsH表达分析检测试剂盒（货号A10067为橙黄色荧光，货号A10068为红色荧光）对蛋白质进行共价标记，不会影响蛋白质在凝胶中的迁移。

电泳期间，可将SYPRO Orange或SYPRO Red蛋白质凝胶染料稀释5000倍并加入到阴极（上层）缓冲液槽中，对蛋白质进行染色而不影响迁移。这样做可能引起的问题是，染料与SDS的相互作用可能导致凝胶产生背景荧光。在电泳后使用7.5%乙酸脱色15–60分钟，可降低这种背景荧光。这种染色方法还会导致蛋白质灵敏度低于标准的后染色方法、在凝胶成像前需要相同的时间以及污染电泳装置。

If I increase the final concentration of my SYPRO Orange or SYPRO Red staining solution above 1X, can I increase the signal of my stained proteins?

No. Dye concentrations higher than 1X do not give better detection. Instead, background fluorescence increases and, as the dye concentration increases, the dye becomes self-quenching and the signal actually decreases.

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

Can I dry my SYPRO Ruby, SYPRO Orange, or SYPRO Red stained gels?

Gels stained with SYPRO dyes can be dried between sheets of cellophane, although there is sometimes a slight decrease in sensitivity. If the gels are dried onto paper, the light will scatter and the sensitivity will decrease. Other plastics are not recommended, as the plastic typically used is not transparent to UV light.

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

Can I pre-stain proteins with SYPRO Ruby, SYPRO Orange or SYPRO Red Protein Gel Stains and then run them through a gel?

No. Loading solutions contain so much SDS that SYPRO Ruby, SYPRO Orange and SYPRO Red dyes simply localize in the free SDS and bind very little of the proteins. Proteins can be covalently pre-labeled with ATTO-TAG CBQCA (Cat. No. A2333), DDAO succinimidyl ester (Cat. No. C34553) or TAMRA-succinimidyl ester (Cat. No. C2211) dyes, or the TC-FLAsH Expression Analysis Detection Kits (Cat. No. A10067 for orange fluorescence, Cat. No. A10068 for red fluorescence) prior to electrophoresis without affecting protein migration through the gel.

SYPRO Orange or SYPRO Red Protein Gel Stain can be diluted 5000-fold into the cathode (upper) buffer tank to stain proteins during electrophoresis without affecting migration. The problem with doing this is that there is considerable background fluorescence in the gels from the dye interacting with SDS. This can be reduced after electrophoresis by destaining in 7.5% acetic acid for 15-60 minutes. This method also results in poorer protein sensitivity than the standard post-staining method, requires the same amount of time before the gel can be imaged, and contaminates the electrophoresis apparatus.

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