Search
Search
View additional product information for CellTrace™ BODIPY™ TR Methyl Ester (Lipophilic Counterstain For GFP) - FAQs (C34556)
14 product FAQs found
我们提供小份的CellTrace试剂，强烈建议您丢弃任何未用的DMSO染料储液。CellTrace试剂有乙酰酯基团，可以覆盖染料的电荷使得它们可以透过细胞，并且琥珀酰亚胺酯胺反应分子允许共价结合到细胞组件，可长时间滞留。如果储存环境中中含有水，乙酰酯和琥珀酰亚胺酯都易于水解。DMSO是吸水性的，因此极易从大气中吸收水。如果您必须要储存自己的染料储备液，需要使用高质量的无水DMSO储液（且不能经常打开），并将小瓶密封放置于密封的含有干燥剂的地方，尽可能保持DMSO/染料储液干燥。
以下为有助于实现均一染色，明亮、清晰的亲代增殖峰的建议：
•临使用前，使用试剂盒附带的DMSO或高质量无水DMSO溶解CellTrace染料储备液，获得最佳的反应活性和细胞透性。
•在PBS或其他无胺、无蛋白生理学缓冲液染色，不要在培养基中染色。
•从单细胞悬液开始染色，并在染色过程中轻轻摇动细胞。
•通过在冷的培养基中孵育细胞5分钟，快速去除未结合的染料，之后用预热培养基洗涤洗细胞两遍。
•在检测体系中引入死细胞染料，并仅对活细胞设门限。
•尽可能多地分析每个样品中的细胞。
•流体动力聚焦细胞流式细胞仪使用低的流动速率。
•CellTrace染料最好的染色浓度通常介于1-10 µM之间，但针对某些类型细胞的最佳浓度会存在差异。观察不同稀释度的染色剂的染色结果，确定您的细胞最适宜的染色浓度。
•某些类型的细胞吸收染料后的染色强度分布范围较广。如果您的细胞也是这种情况，则需要首先分选染色的未刺激亲本细胞，从而选择出一个窄的峰分布。
我们提供小份的CellTrace试剂并强烈建议丢弃任何未用的DMSO染料储备液。CellTrace试剂有二醋酸基团可以封闭染料的电荷，使得它们可以透过细胞并且琥珀酰亚胺酯胺反应基团可允许长时间滞留。如果储存环境中含有水，二醋酸盐和琥珀酰亚胺酯都易于水解。DMSO是吸水性的因此极易从大气中吸收水。如果您必须要储存您的染料储备液，您需要使用高质量的无水DMSO储备液，不能经常打开且将小瓶密封放置于密封的含有干燥剂的地方来保持DMSO/染料储备液尽可能的干燥。-20°C保存。在短时间内尽快使用。
CellTrace细胞增殖试剂是一种细胞透过型染料，胞内酯酶可将其分解产生高荧光化合物并可以共价地结合到细胞内的胺类上，将染料附着于多种细胞内组分并产生稳定的信号。这些试剂毒性很小并对多种细胞增殖活性的影响很小。
用CellTrace试剂绘制出好的传代曲线的关键是在开始时均匀标记细胞，使它们在标记零点有紧密的变异系数（CV）。如果峰太宽，迭代相互重叠，一系列峰将成为一个驼峰。由于染色程度与细胞大小成正比，可以从统一的细胞群（而非淋巴细胞和粒细胞等混合的细胞群）开始标记，实现均匀的标记。细胞标记时间很短，所以您需要预先稀释染料，迅速混入你的细胞。请确保您的细胞未沉积在试管底部。实现这一点最简单的方法就是准备一份2X染色液（1×= 1-10μM），在一半体积（无血清或BSA）培养基中重悬细胞。将染料添加到细胞中并倒置几次以混合。染色过程中轻轻晃动细胞。染料孵育结束后（20分钟，37℃）即添加血清或BSA（至少1％），以清除任何残留的未反应染料。离心细胞，洗涤一遍，然后在完全培养基中重悬。经10-20分钟的脱酯化孵育后，细胞即可用于你们的实验。一定要确保有一个零点对照，因为您需要知道细胞第一次传代的时间。
•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。
We provide the CellTrace reagents in small aliquots and strongly recommend discarding any unused DMSO/dye stocks. The CellTrace reagents have acetyl groups to cap the charges on the dyes to make them cell permeant, and succinimidyl ester amine-reactive moiety to allow for covalent attachment to cellular components for long-term retention. Both acetyl groups and succinimidyl esters will readily hydrolyze if any water is present during storage. DMSO is hygroscopic and thus readily absorbs water from the atmosphere. If you must store your dye stocks, you will need to use a good quality, anhydrous DMSO stock that has not been opened often and store the vial within an air-tight container containing some desiccant to keep the DMSO/dye stock solution anhydrous during storage.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Here are some tips to obtain uniform staining and a bright, unstimulated parent generation peak:
Dissolve the CellTrace dye stock immediately before use in the DMSO provided in the kit or in good quality, anhydrous DMSO to obtain the best reactivity and cell permeability.
Stain in PBS or other amine-free, protein-free physiological buffer. Do not stain in medium.
Start with a single-cell suspension and gently agitate the cells during staining.
Quickly remove the unbound dye by incubating the cells in ice-cold media for 5 minutes and then wash twice more with pre-warmed media.
Include a dead-cell stain in the assay and gate only on live cells.
Analyze as many cells as possible from each sample.
Use a low flow rate for analysis on hydrodynamic focusing cytometers.
A good staining concentration for the CellTrace dyes is generally within 1-10 µM, but the optimal concentration for a particular cell type will vary. Observe your cells in a stain dilution series to determine the optimal concentration for your cells.
Some cell types may take up dye with a broad staining intensity distribution. If this is the case for your cells, then you will need to do an initial sort of the stained, unstimulated parent cells to select for a narrow peak distribution.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
We provide the CellTrace reagents in small aliquots and strongly recommend discarding any unused DMSO dye stocks. Some of the CellTrace reagents have diacetate groups to cap the charges on the dyes to make them cell permeant, and some have succinimidyl ester amine-reactive groups for long-term cellular retention. Both diacetates and succinimidyl esters will readily hydrolyze if any water is present during storage. DMSO is very hygroscopic and thus readily absorbs water from the atmosphere. We do not recommend storing stock solutions of CellTrace reagents because storage of the product in solution will inevitably lead to partial or complete loss of reactivity.
CellTrace Cell Proliferation reagents are all cell-permeant dyes that are cleaved by intracellular esterases to yield highly fluorescent compounds that also covalently bind to cellular amines, attaching the dye to various cellular components and providing a very stable signal. These reagents show little cytotoxicity with minimal observed effects on the proliferative ability of many cells.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The key to good generational profiles with CellTrace reagents is starting with cells that are evenly labeled so that they have a tight coefficient of variance (CV) when run at time zero after labeling. If the peak is too broad, the generations will overlap each other and the series of peaks will become a hump. Even labeling can be achieved by starting with a uniform cell population (not a mixture of lymphocytes and granulocytes for example) as staining will be proportional to cell size. Cells are labeled rapidly, so you want to pre-dilute the dye and mix it into your cells rapidly. Be sure that the cells are not sitting in a clump in the bottom of your tube. The easiest way to do this is to make a 2x dye solution (1x = 1-10 µM) and resuspend your cells in a half volume of medium (no serum or BSA). Add the dye to the cells and invert a few times to mix. Gently agitate the cells during staining. Once the dye incubation is over (20 min, 37 degrees C), add serum or BSA (at least 1%) to scavenge any remaining unreacted dye. Spin down cells, wash 1x, and resuspend in complete medium. After a 10-20 min incubation to undergo de-esterification, cells are ready to be set up for whatever treatment you are planning. Be sure to keep a time zero control as you need to know where the first generation ran.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
-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.