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View additional product information for Click-iT™ TUNEL Colorimetric IHC Detection Kit - FAQs (C10625)
16 product FAQs found
可以，额外的末端脱氧核苷酸转移酶（rTdT）可以购买。货号为10533-065或10533-075。额外的TdT反应缓冲液可以购买，货号为16314-015。
我们没有在3D培养体系中使用过EdU TUNEL检测调亡，但是因为这种试剂适用于体内细胞标记，所以它也有希望可以标记3D培养体系中的细胞。文献中有大量关于在3D培养体系中使用这种产品报道；此处有一些引证：
Lei Y, Schaffer DV (2013) A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation.Proc Natl Acad Sci U S A 110:E5039–E5048.
Derda R, Laromaine A, Mammoto A et al.(2009) Paper-supported 3D cell culture for tissue-based bioassays.Proc Natl Acad Sci U S A 106:18457–18462.
Robertson FM, Ogasawara MA, Ye Z et al.(2010) Imaging and Analysis of 3D Tumor Spheroids Enriched for a Cancer Stem Cell Phenotype.J Biomol Screen 15:820–829.
大部分组织样品需要充分消化15分钟。最佳的培养时间根据组织类型和厚度而不同。我们已经观测到脑组织比其他组织需要更长的蛋白酶处理时间。
我们在5–20 µM 厚度的老鼠肠、肾脏、肝脏、心脏和结肠FFPE切片中验证了Click-iT Plus TUNEL for In Situ Apoptosis Detection实验
初代Click-iT TUNEL实验试剂盒针对细胞培养样品优化，也可用于组织样品。我们建议对组织样品使用Click-iT Plus TUNEL for In Situ Apoptosis Detection实验试剂盒；用于组织的Click-iT Plus TUNEL for In Situ Apoptosis Detection实验试剂盒的实验方案针对组织样本优化。通过改进方案可以提高TdT酶进入多层细胞组织样品的效果。另外，相比于Click-iT Plus TUNEL试剂盒，我们发现初代Click-iT TUNEL试剂盒可能会在组织样品出现高度非特异性结合和点状染色。有一个增加组织透过性较好的方法是用蛋白酶K消化然后甲醛中再固定样品以取代去污剂透化作用步骤。Click-iT Plus TUNEL试剂盒手册第三部分的组织固定和透化作用方案可用于初代Click-iT TUNEL实验处理组织样品。胃蛋白酶和其他的蛋白水解酶同时也可用于提高组织透过性。
•可测定来自单个细胞的数据。
•可从大量细胞中获得数据，产生细胞群的丰富统计学分析结果。
•由于可测定单个细胞，能够揭示种群的异质性。
•支持多重检测，可鉴定小型亚群。
•可以快速分析数以千计的细胞。
•非常适合血液样本和其他悬浮细胞。
•数据获取后，可以多次重复分析。
•流式细胞仪文件（FCS）可以归档。
可进行多种应用，包括免疫分型、细胞周期分析、凋亡检测（如膜联蛋白V染色检测实验）、CellEvent Caspase-3/7检测、TUNEL检测、细胞活性检测、增殖检测（如CellTrace 检测和Click-iT EdU检测）、MitoProbe检测法测定线粒体电势、利用计数微球进行细胞计数。
Yes, but we have not optimized its use with this kit.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Yes, other colorimetric stains can be used.
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Yes, additional Terminal Deoxynucleotidyl Transferase (rTdT) can be purchased as Cat. No. 10533-065 or 10533-075. Additional TdT reaction buffer can be purchased as Cat. No. 16314-015.
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We have not validated the use of EdU TUNEL for apoptosis detection in 3D culture systems, but as this reagent is compatible for labeling cells in vivo, it is also expected to label cells in 3D culture systems. There are a number of reports in the literature that use this product in 3D culture systems; here are some citations:
Lei Y, Schaffer DV (2013) A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation. Proc Natl Acad Sci U S A 110:E5039-E5048.
Derda R, Laromaine A, Mammoto A et al. (2009) Paper-supported 3D cell culture for tissue-based bioassays. Proc Natl Acad Sci U S A 106:18457-18462.
Robertson FM, Ogasawara MA, Ye Z et al. (2010) Imaging and Analysis of 3D Tumor Spheroids Enriched for a Cancer Stem Cell Phenotype. J Biomol Screen 15:820-829.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Most tissue samples will be adequately digested in 15 minutes. The optimal incubation time will vary depending on tissue type and thickness. We have observed that brain tissue needs longer proteinase K treatment than other tissues tested.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
We have validated the Click-iT Plus TUNEL for In Situ Apoptosis Detection assay on 5-20 µM thick FFPE sections of mouse intestine, kidney, liver, heart, and colon.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The original Click-iT TUNEL assay kits were optimized for cell culture samples and may also be used on tissue samples. We recommend using the Click-iT Plus TUNEL for In Situ Apoptosis Detection assay kits on tissue samples; the protocols for the Click-iT Plus TUNEL for In Situ Apoptosis Detection assay kits have been optimized for tissue. The protocol was modified to improve accessibility of the TdT enzyme into the multiple cell layers of tissue samples. In addition, we found that the original Click-iT TUNEL kit may show higher non-specific binding and punctate staining in tissues compared to the Click-iT Plus TUNEL kit. One method that works well to increase permeability for tissues is to replace the detergent permeabilization step with proteinase K digestion and then refix the sample in formaldehye. The Tissue Fixation and Permeabilization protocol in section 3 of the Click-iT Plus TUNEL kit manual can be followed for tissue samples using the original Click-iT TUNEL assay. Pepsin and other proteolytic enzymes can also be used to improve permeability.
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-Measures data from single cells.
-Data are obtained for a large number of cells, generating a rich statistical analysis of cell populations.
-Because single cells are measured, it will reveal heterogeneity within a population.
-With the ability to multiplex, small sub-populations can be identified.
-Thousands of cells can be analyzed rapidly.
-It is ideally suited for blood samples and other cells in suspension.
-Data can be re-analyzed multiple times after acquisition.
-Flow cytometry files (FCS) can be archived.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
There are several applications, some of which include immunophenotyping, cell cycle analysis, apoptosis assays such as annexin V staining, CellEvent Caspase-3/7 assay, and TUNEL assay, cell viability, proliferation assays such as CellTrace assay and Click-iT EdU assay, measurements of mitochondrial potential with MitoProbe assays, and cell counting using counting beads.
Find additional tips, troubleshooting help, and resources within our Flow Cytometry Support Center.