| Product Name | Calcein, AM |
| CAS | 148504-34-1 |
| Size | 1 mg |
| Catalog # | AS-89201 |
| US$ | $143 |
| Purity | ≥95% by HPLC |
Calcein, AM is a cell-permeant and non-fluorescent compound that is widely used for determining cell viability.In live cells the non-fluorescent calcein, AM is hydrolyzed by intracellular esterases into the strongly green fluorescent anion calcein. The fluorescent calcein is well-retained in the cytoplasm in live cells.Hela cells stained with AnaSpecs Calcein AM fluorescent dye (Cat# AS-89201, AS-89202, AS-89203 and AS-89204). MCF-7 cells grown in 8-well slide chamber for 2 days. Cells were washed twice before adding 1 uM Calcein AM (AnaSpec cat# AS-89202). Images acquired every 3 seconds and total elapsed time on video is 12 min.
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| Images | ![]() |
| Detailed Information | |
| Storage | -20°C desiccated and protected from light |
| Molecular Weight | 994.857 |
| Molecular Formula | C46H46N2O23 |
| Spectral Properties | Abs/Em = 494/517 nm |
| Solvent System | DMSO |
| Product Citations | Graham, DM. et al. (2012). Spatial manipulation of cells and organelles using single electrode dielectrophoresis. Int J of Life Sci Meth 52 , 39-43Kim, H. et al. (2011). Human macrophage ATP7A is localized in the trans-golgi apparatus, controls intracellular copper levels, and mediates macrophage responses to dermal wounds. Inflammation 10.1007/s10753-011-9302-z.Decaris, ML. et al. (2010). Design of experiments approach to engineer cell-secreted matrices for directing osteogenic differentiation. Annals Biomed Eng 39, 1174.Hernlem, B. et al. (2010). Dual fluorochrome flow cytometric assessment of yeast viability. Current Microbiol 61, 57.Miura, Y. et al. (2010). Change of morphological and functional characteristics of retinal pigment epithelium cells during cultivation of retinal pigment epithelium-choroid perfusion tissue culture. Opthalmic Res 43, 122.Zaveri, T. et al. (2010). Contributions of surface topography and cytotoxicity to the macrophage response to zinc oxide nanorods. Biomater 31, 2999.Decaris, ML. et al. (2009). Influence of the oxygen microenvironment on the proangiogenic potential of human endothelial colony forming cells. Angiogenesis 12, 303.Delaunay, JL. et al. (2009). A missense mutation in ABCB4 gene involved in progressive familial intrahepatic cholestasis type 3 leads to a folding defect that can be rescued by low temperature. Hepatology 49, 1218. Zhang, K-H. et al. (2009). Ferritin heavy chainmediated iron homeostasis and subsequent increased reactive oxygen species production are essential for epithelial-mesenchymal transition. Cancer Res. 10.1158/0008-5472.CAN-09-0112. |
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1、 缓冲溶液对反应物的测定没有干扰
2、缓冲组分的浓度为1:1
3、有足够的缓冲容量
4、缓冲溶液的PH应在所需范围内
5、组成缓冲溶液的弱碱PKB和弱酸PKA应接近或等于所需的POH值或PH值(PH+POH=14)
配制
只要知道缓冲对的PH值,和要配制的缓冲液的pH值(及要求的缓冲液总浓度),就能按公式计算[盐]和[酸]的量。这个算法涉及对数换算,较麻烦,前人为减少后人的计算麻烦,已为我们总结出pH值与缓冲液对离子用量的关系并列出了表格。只要我们知道要配制的缓冲液的pH,经查表便可计算出所用缓冲剂的比例和用量。例如配制500nmpH5.8浓度为0.1M磷酸缓冲液。
经查表知pH5.8浓度为0.2M Na2HPO48.0毫升,而0.2M Na2HPO492.0毫升。依此可推论出配制100ml0.1M的磷酸缓冲液需要0.1M Na2HPO48.0毫升,而0.1M Na2HPO4需要92.0毫升。
计算好后,按计算结果准确称好固态化学成分,放于烧杯中,加少量蒸馏水溶解,转移入50ml容量瓶,加蒸馏水至刻度,摇匀,就能得到所需的缓冲液。
各种缓冲溶液的配制,均按表格按比例混合,某些试剂,必须标定配成准确浓度才能进行,如醋酸、氢氧化钠等。另外,所有缓冲溶剂的配制计量都能从以上的算式准确获得。
TBST中含有Tris-Hcl,NaCl,Tween20这三种物质,是做WESTERNBLOT中常用的一种缓冲液。
TBST缓冲液的配制
1000ml×TBST的配置
先称量NaCl40g,倒入烧杯中,加DDW蒸馏水400ml,再称量NaCl47.6g,倒入刚才的那个烧杯中(PS:由于NaCl的量太多,一次称量不方便,所以分两次称量,且易于溶解)。往烧杯中加入Tris—HCl缓冲液100ml,最后加(吐温20)5ml,转入1000ml容量瓶中,在定容,转移即可。
TBST缓冲液的应用:
1.主要用于免疫组化和原位杂交,酶联免疫等实验中,清洗免疫印。
2.迹膜;
注意事项:
1.TBST缓冲液,PH7.2-7.5;
2.颜色为无色透明液体;
3.为了您的安全和健康,请穿实验服并戴防护手套操作;



