Biotinylated liposomes can be conjugated non-covalently with (strept)avidin through either direct interaction with the protein/antibody conjugated to (strept)avidin or by coupling with other biotinylated proteins using (strept)avidin as a bridging molecule. Both avidin and (strept)avidin form strong non-covalent bond with biotin. The high resistance to breakdown makes them very useful in bioconjugate chemistry. However, (strept)avidin has replaced avidin in most bioconjugation applications due to its enhanced properties. NeutrAvidin (ThermoFisher) is a modified avidin without negative properties. It performs much better than original avidin and sometimes (strept)avidin.
In order to exploit the high-affinity interaction of biotin with (strept)avidin, a two-step “sandwich” protocol (Method A) has been developed for the preparation of targeted immunoliposomes. In this methodology, (strept)avidin is first attached to biotinylated liposomes, then a biotin-modified protein/antibody is introduced into the biotinylated (strept)avidin-labeled liposomes. This non-covalent approach is rapid, extremely versatile and applicable to numerous targeting ligands of interest with respect to in vitro and in vivo applications. Alternatively, instead of forming a (strept)avidin bridge, (strept)avidin molecule can also be covalently conjugated to antibody or ligand (Method B) and non-covalently bound to liposomes containing biotin on surface in order to form immunoliposomes.
ImmunoFluor™-Biotin is a Non-PEGylated product. For other reactive (PEGylated and non-PEGyalated products) biotinylated liposomes and also ImmunoFluor™ products suitable for other types 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？
