Lumio™ Green 检测试剂盒
Lumio™ Green 检测试剂盒
Invitrogen™

Lumio™ Green 检测试剂盒

The Lumio™ Green Detection Kit is a sensitive and highly specific kit for labeling Lumio™ fusion proteins prior to electrophoresis.了解更多信息
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货号数量
LC60901 kit
货号 LC6090
价格(CNY)
8,415.00
Each
添加至购物车
数量:
1 kit
请求批量或定制报价
价格(CNY)
8,415.00
Each
添加至购物车
The Lumio™ Green Detection Kit is a sensitive and highly specific kit for labeling Lumio™ fusion proteins prior to electrophoresis. The Lumio™ Green Kit enables immediate visualization of Lumio™ fusion protein bands directly in a polyacrylamide gel. Using the Lumio™ Green Detection Kit you'll:

• Label proteins fused to a Lumio™ sequence
• Visualize results immediately after electrophoresis—eliminate gel staining and western blotting
• Detect nanogram levels of protein with a UV transilluminator, equipped with a standard camera (Figure 1), or a laser-based scanner
The Lumio™ Green Detection Reagent, included in the Lumio™ Green Detection Kit, can also be used for real-time detection of protein synthesis during in vitro transcription-translation reactions.

Immediate and Effortless Detection
To detect using the Lumio™ Green Detection Kit simply:
1. Add the Lumio™ Green Detection Reagent and the optimized Lumio™ Sample Buffer to your protein sample and heat for 10 minutes at 70°C.
2. Add the Lumio™ In-Gel Detection Enhancer to your sample mix and incubate for 5 minutes at room temperature.
3. Load and electrophorese your protein gel.
4. Visualize your Lumio™ fusion protein bands with a UV transilluminator, equipped with a standard camera, or laser-based scanner.
5. After documenting your Lumio™ Green signal, total protein staining can be performed.
仅供科研使用。不可用于诊断程序。
规格
检测定位凝胶内检测
检测方法荧光
产品线Lumio
产品类型预制凝胶融合蛋白标记试剂盒
数量1 kit
有效期每 6 个月
运输条件干冰
靶标分子蛋白质(带有 Lumio™ 标记)、蛋白质(带有 TC 标记)
标签或染料Lumio Green
Unit SizeEach
内容与储存
Lumio™ 绿色检测试剂盒包含一瓶 Lumio™ 绿色检测试剂、五瓶 Lumio™ 凝胶样品缓冲液,以及一瓶 Lumio™ 胶内检测增强剂。储存于 -20°C 下。如果适当储存,所有组分可保持稳定 6 个月。

常见问题解答 (FAQ)

Lumio试剂具有透膜能力么?

Lumio试剂具有疏水性,因此很容易穿过细胞膜。无需通过透化处理来帮助该试剂进入细胞内部。 

在使用Lumio标记哺乳动物细胞的过程中,BSA会有背景干扰吗?

哺乳动物细胞培养基中的血清蛋白(如BSA,66KD)可能会与Lumio试剂发生交叉反应,从而形成非特异性的条带。吸去细胞培养基并在收获细胞后使用PBS润洗哺乳动物细胞3-4遍,有助于减少BSA的非特异性结合。 

Lumio Red(红)与Green(绿)试剂对细胞有毒性吗?

我们尚未在细胞中发现用于蛋白检测浓度的Lumio试剂具有任何不良效应。我们也未在使用Lumio Green之后发现细胞形态出现任何不良改变。加入Lumio Red后,我们确实发现细胞发生了某些微小的形态改变,不过这一变化在加入试剂24小时之后恢复。 

Lumio染色与GeneBLAzer检测相比效果如何?GFP作为目的基因的检测手段效果如何?

Lumio染色技术的优势在于可同时兼容体内和体外的蛋白标记操作。在体内标记实验中,向细胞中加入Lumio试剂后即可在荧光显微镜下观察到细胞/蛋白。这一效果与GeneBLAzer检测步骤相似,只是GeneBLAzer是基于扩增报告基因信号的酶促反应。GFP的荧光信号只能在细胞(体内)中检测到,因为需要GFP蛋白的正确折叠。相比GeneBLAzer检测法中的bla蛋白(264个氨基酸,29 kDa)和GFP蛋白(27 kDa)而言,Lumio标签非常之小(6个氨基酸,585Da),因此在很大程度上不会对所融合蛋白的功能造成影响。GFP的劣势在于需要融合一个很大的标签蛋白,而且该检测法并非是基于酶法的报告系统。不同于GeneBLAzer检测法和GFP标签,Lumio-标签蛋白可在Lumio试剂处理细胞裂解液或蛋白之后通过凝胶电泳变得可视化。与Lumio和GFP相比,GeneBLAzer检测法在活体细胞的应用中更为灵敏。GeneBLAzer检测法也不同于Lumio和GFP,这一方法能够实现比率化的读数,因此有助于减少样本间的差异。

我在使用Lumio绿色检测试剂盒时,得到了较高的、不均匀的背景。你们有何建议?

以下是可能原因和解决方案:

•凝胶处理不恰当或成像平台不干净。处理凝胶或凝胶成像时,不要用手直接接触凝胶。为将所有背景荧光降至最低,每次进行凝胶成像前都应使用纸巾清洁成像系统。
•蛋白上样量过多。降低蛋白浓度或减少上样体积。
•非特异性结合。为将非特异性结合降至最低,应使用Lumio胶内检测增强剂。大肠杆菌裂解物中的某些蛋白(SlyD,21 kDa)和哺乳细胞培养基中的牛血清白蛋白(BSA,66 kDa),会与Lumio绿色试剂发生交叉反应,产生非特异性结合。移除细胞培养基,收获细胞后使用PBS洗涤哺乳细胞3-4次,从而使BSA的非特异性结合降至最低。

引用和文献 (4)

引用和文献
Abstract
In vivo oligomerization and raft localization of Ebola virus protein VP40 during vesicular budding.
Authors:Panchal RG, Ruthel G, Kenny TA, Kallstrom GH, Lane D, Badie SS, Li L, Bavari S, Aman MJ,
Journal:Proc Natl Acad Sci U S A
PubMed ID:14673115
The matrix protein VP40 plays a critical role in Ebola virus assembly and budding, a process that utilizes specialized membrane domains known as lipid rafts. Previous studies with purified protein suggest a role for oligomerization of VP40 in this process. Here, we demonstrate VP40 oligomers in lipid rafts of mammalian ... More
Scalable and Versatile Genome Editing Using Linear DNAs with Microhomology to Cas9 Sites in Caenorhabditis elegans.
Authors:Paix A, Wang Y, Smith HE, Lee CY, Calidas D, Lu T, Smith J, Schmidt H, Krause MW, Seydoux G,
Journal:
PubMed ID:25249454
Homology-directed repair (HDR) of double-strand DNA breaks is a promising method for genome editing, but is thought to be less efficient than error-prone nonhomologous end joining in most cell types. We have investigated HDR of double-strand breaks induced by CRISPR-associated protein 9 (Cas9) in Caenorhabditis elegans. We find that HDR ... More
Molecular and functional characterization of IL-15 in rainbow trout Oncorhynchus mykiss: a potent inducer of IFN-gamma expression in spleen leukocytes.
Authors:Wang T, Holland JW, Carrington A, Zou J, Secombes CJ,
Journal:J Immunol
PubMed ID:17641013
IL-15 is a member of the common gamma-chain family of cytokines that possess a heterogeneous repertoire of activities on various cells of the immune system. We report here the first functional characterization of a fish IL-15 in rainbow trout. The trout IL-15 gene is 6-kb long and contains six exons ... More
Tetracysteine genetic tags complexed with biarsenical ligands as a tool for investigating gap junction structure and dynamics.
Authors:Sosinsky GE, Gaietta GM, Hand G, Deerinck TJ, Han A, Mackey M, Adams SR, Bouwer J, Tsien RY, Ellisman MH,
Journal:Cell Commun Adhes
PubMed ID:14681013
Gap junctions (GJ) are defined as contact regions between two adjacent cells containing tens to thousands of closely packed membrane channels. Cells dynamically modulate communication through GJ by regulating the synthesis, transport and turnover of these channels. Previously, we engineered a recombinant connexin43 (Cx43) by genetically appending a small tetracysteine ... More