Product Name | SensoLyte® Red Glucocerebrosidase (GBA) Assay Kit *Fluorimetric*NEW |
Size | 1 kit |
Catalog # | AS-72259 |
US$ | $480 |
Glucocerebrosidase/GBA (also called acid β-glucosidase, Glucosylceramidase,) is a lysosomal enzyme responsible for the breakdown of glucocerebroside releasing glucose and ceramide. Deficiency of this enzyme due to genetic mutations leads to accumulation of glucocerebroside and development of lysosomal storage disease, known as Gaucher disease (GD). Mutations in the glucocerebrosidase (GBA1) gene are also associated with increased risk for Parkinson disease and related disorders. It has been hypothesized that GBA, when not available to clear out proteins like alpha-synuclein, results in the accumulation of the proteins thereby contributing to Parkinsons disease. The SensoLyte® Red Glucocerebrosidase (GBA) Assay Kit detects GBA activity by using a highly sensitive fluorogenic substrate. The GBA substrate provided in the kit releases the red fluorescent dye resorufin upon Glucocerebrosidase cleavage with absorption/emission maxima at 570 nm/610 nm. This assay kit can be used to detect enzyme activity in purified enzyme preparations and compound screening. The Red Glucocerebrosidase (GBA) Assay is one-step homogenous reaction, which does not require the additional step of dispensing stop solution after the incubation, thereby suitable for HTS.The long wavelength fluorescence resorufin is also less interfered by the autofluorescence of components in biological samples and test compounds. Related Product (ex/em=365nm/445nm)SensoLyte® Blue Glucocerebrosidase (GBA) Assay Kit *Fluorimetric*Fig 1.Inhibition of GBA activity by Isofagomine as measured withSensoLyte®Blue Glucocerebrosidase Assay Kit. | |
Detailed Information | DatasheetMaterial Safety Data Sheets (MSDS) |
References |
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(2)荧光标记法 : 使用二乙酸荧光素(FDP)、碘化丙啶(PI)或异硫氰酸荧光素钠标记的荧光染料与细胞共孵育,用流式细胞仪检测荧光染色阳性细胞的比率。此法其实是(1)法的“荧光”版,但其在灵敏性和准确性方面明显要优于后者。
(3)硝酸镧(La)示踪法: 在正常的生物组织中镧微粒可沉积于细胞间隙,但不能穿过具有1~ 2nm 微小间隙的细胞膜性结构(包括细胞膜和细胞器膜),也不能穿过细胞间的紧密连接。在膜性结构通透性增高时, 镧微粒则可进入细胞、细胞器和紧密连接内, 并在电镜下显示, 镧盐标记技术被认为是一种有效的监测细胞膜通透性变化的标记技术。
(4)LDH释放法: 在正常情况下,细胞内大分子物质LDH 是不能通过细胞膜的, 但在细胞膜受损伤而通透性增加时,可通过受损的细胞膜释放出来。LDH 能较好地反映细胞膜损伤程度。类似的还有检测细胞外K+的漏出率等。
有几个疑问
1:荧光标记到细胞是标记到细胞表面还是细胞质内?
2:荧光应该随着细胞的分化和增殖逐渐消失?是不是分化增殖越快,荧光消失速度越快?
3:有哪些容易操作,成本便宜的荧光物质?
谢谢各位战友
比如你用的CD1a-FITC(如果是鼠单抗IgG1,那对照抗体就要用相同物种的非特异性IgG1-FITC)。注意浓度要相同。一般提供抗体的公司BD,santa cruz等有提供的。其他就按照说明书的推荐浓度和孵育时间。