EGFP-rProtein APDF
Catalog Number:
E1GA1-020Amount:
200μg/200μlBackground:
The unique, high quality recombinant EGFP-rProtein A are produced in E. coli, and purified using multiple chromatographic steps (not affinity purified with human IgG). EGFP-rProtein A bind to most mammalian immunoglobulins in their Fc binding domains and therefore are ideal for immunofluorescence staining. The genetically engineered protein was expressed and purified from transformed E. coli containing only the IgG binding regions and have 6X His-tag on their N-terminus. Other domains such as cell wall binding region, albumin binding region and other non-specific binding regions have been eliminated from all these proteins to ensure the maximum specific IgG binding and maximal fluorescence.Form of Antibody:
Liquid. The product is 1 mg/ml in 10 mM Phosphate buffered saline, pH 7.4 containing, 10% glycerol and 0.1% sodium azide as preservative.Source:
Recombinant expressed in E. coliPurity:
>98% by SDS-PAGE and HPLCEndotoxin:
< 0.05="">IgG Binding:
>95%Bioburden:
No organisms detectedSpecificity/Sensitivity:
Under optimal conditions, 1 mg EGFP-rProtein A will bind approximately 5 mg IgG. Optimal binding of EGFP-rProtein A to antibodies occurs at pH 5.0 to 8.0 and can be eluted over a pH range of 2.5 to 3.0.Applications:
The protein is suitable as control reagent for EGFP expression studies or as labeling reagent. Applications include: Flow cytometers, fluorescence microscopy, and microinjection, antibody purification, etc. at 1:100 dilution. Maximum absorption wavelength at 488 nm and emission wavelength at 507 nm.Storage/Stability:
Store at -20°C. -80°C for long-term storage. Avoid exposure to light. Avoid repeated freezing and thawing.293T cells were stained with mouse anti-beta-tubulin antibody (E1C601) and detected with EGFP-rProtein A (Green).
Price:
160$

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有没有朋友使用碧云天的细胞衰老β-半乳糖苷酶染色试剂盒做过细胞衰老,本人小白,求相关细胞处理的过程及数据分析方法。贴壁细胞六孔板需要接种多少,染色后怎样计数进行数据处理。谢谢啦!
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
的抗体不容易染色,哪为同仁有这方面的经验?另外基因公司卖的cellsignalingtechnology的抗体要1875元,而中山公司卖的这个抗体只有800多,其中差距在哪里?请赐教
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?

