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Glutaraldehyde, EM Grade, 8%


Company: Polysciences
Catalog#: 111-30-8
Other protocol()

Differentiation of Naturally Produced Extracellular Membrane Vesicles from Lipid Aggregation by Glucuronoxylomannan Immunogold Transmission Electron Microscopy in Bacillus subtilis
[Abstract]  Recently, membrane vesicle (MV) production was described in Gram-positive bacteria, which harbor a variety of components such as toxins, antibiotic resistance proteins, proteases, DNA, and immune modulators. Free lipids have the ability to form micelles, thus it is important to rule out spontaneous association of lipids into vesicle-like structures and rather, that MVs are produced naturally by a metabolically active cell. Here, we describe a protocol utilizing the polysaccharide, glucuronoxylomannan (GXM) from Cryptococcus neoformans (C. neoformans) as a marker to differentiate naturally produced MVs from vesicles that form spontaneously in the Gram-positive model organism, Bacillus subtilis (B. subtilis). MVs are purified from bacterial cultures grown ... [摘要]  最近,在革兰氏阳性细菌中描述了膜囊泡(MV)产生,其含有多种组分,例如毒素,抗生素抗性蛋白,蛋白酶,DNA和免疫调节剂。游离脂质具有形成胶束的能力,因此排除脂质自发缔合成囊泡样结构是重要的,并且更确切地说,MV由代谢活性细胞天然产生。在这里,我们描述了利用来自新隐孢子虫的多糖葡萄糖醛酸甘露聚糖(GXM)( C。新型隐球菌)作为标记物来区分天然产生的MV与自发形成的囊泡革兰氏阳性模式生物,枯草芽孢杆菌(<枯草芽孢杆菌)。从在GXM存在下生长的细菌培养物中纯化MV;由细胞天然产生的MV在管腔中不含有GXM,而在培养基中形成的囊泡结构可包封GXM,并且这可通过免疫金属透射电子显微镜可视化。

Colon Tissue Immunoelectron Microscopy
[Abstract]  The method described here is intended to study intracellular localization of proteins in colon cells. This protocol was used to localize IRE1β in the endoplasmic reticulum membrane. We used anti-IRE1β antibody raised in guinea pig for this purpose. We also studied the location of BiP (also known as GRP78), with the antibody raised in rabbit. Both antibodies used with appropriate gold particle-conjugated secondary antibodies gave good results. Primary mouse antibodies are not recommended because secondary anti-mouse antibodies also react with the internal mouse epitopes. [摘要]  本文所述的方法旨在研究结肠细胞中蛋白质的细胞内定位。 该方案用于定位IRE1β在内质网膜中。 我们使用在豚鼠中产生的抗IRE1β抗体用于此目的。 我们还研究了在兔中产生的抗体的BiP(也称为GRP78)的位置。 与合适的金颗粒缀合的二抗一起使用的两种抗体都给出了良好的结果。 不推荐初级小鼠抗体,因为次级抗小鼠抗体也与内部小鼠表位反应。