Numerous techniques have been developed to prepare immunoliposomes based on the nucleophilic reactivity of free amine groups of proteins or peptides. However, the most common, versatile and straightforward activation chemistry for creating reactive acylating reagents and labeling peptides/proteins is to form NHS ester with primary amines. A single step nucleophilic substitution reaction between NHS ester derivative and alpha amines at the N terminal or the beta amines of lysine side chains leads to the formation of a stable amide bond.
N-Hydroxysuccinimide (NHS) esters of DSPE-PEG-NHS liposomes react with the primary amine groups on the peptides, proteins, antibodies or other functional ligands for targeted drug delivery applications. The nucleophilic attack of the amine groups on the NHS-activated carbonyl group of the DSPE-PEG-NHS results in the elimination of the NHS group and formation of amide linkage.
ImmunoFluor™-NHS is a PEGylated product. For other amine reactive (PEGylated and non-PEGyalated) products and also ImmunoFluor™ products suitable for other types of conjugation methods see here.
ebiomall.com
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Bone Mesenchymal Stem Cells 作为一个细胞群体，还没有发现有特定细胞表面marker. 对于那些可以代表自我更新和分化的marker, 也不清楚到底要发现哪一个的表达才能确定该细胞就是BMSC。
目前常用的方法，就是采用培养，colony-forming unit-fibroblasts (CFU-F)这个方法。一般BMSC可以24-48小时贴壁。
流式细胞计数，比如STRO-1，但是一般认为STRO-1阳性的细胞更趋向于造血干细胞，和BMSC简单区别还不是很清楚。
这里有个培养分化的产品
http://www.rndsystems.com/pdf/SC020.pdf
GlucosestarvationcausestranslocationofAMPKβ2tothelysosomeinHEK-293cellsthatisdependentonN-myristoylation.Theexperimentwasperformedinβ2KOcellsasinFig.1c,exceptthatthelysosomalMarkerLAMP1(taggedwithRFP)wasco-expressedwiththewild-typeormutantAMPKβ2.Upperpanelsshowmergedimagesstainedblue(4′,6-diamidino-2-phenylindole(DAPI),nuclei),red(LAMP1,lysosomes)andgreen(AMPKβ2,detectedusingantibodyvalidatedine),incellsincubatedwithorwithoutglucosefor20 min.Lowersmallpanelsaremagnificationsoftheareasindicatedbydashedboxesintheupperpanels,showing(LtoR)redandgreenchannelsandmergedimages.
下面的这段话是图注，图注的意思我明白，但是我想知道merge后的图看什么颜色的荧光，蓝色是细胞核，红色是lysosome(位于胞质），绿色是AMPKβ2，该实验是想观察AMPKβ2是否转位到lysosome上了，如果确实发生了AMPKβ2转位到lysosome上，那么merge后是红色与绿色融合在一起，是吗？融合在一起发什么颜色的光了？
