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HBSS, no calcium, no magnesium, no phenol red

HBSS,无钙,无镁,无酚红

Company: Thermo Fisher Scientific
Catalog#: 14175095
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VLA-4 Affinity Assay for Murine Bone Marrow-derived Hematopoietic Stem Cells
Author:
Date:
2017-02-20
[Abstract]  Hematopoietic stem cells (HSCs) are defined by their functional ability to self-renew and to differentiate into all blood cell lineages. The majority of HSC reside in specific anatomical locations in the bone marrow (BM) microenvironment, in a quiescent non motile mode. Adhesion interactions between HSCs and their supporting BM microenvironment cells are critical for maintaining stem cell quiescence and protection from DNA damaging agents to prevent hematology failure and death. Multiple signaling proteins play a role in controlling retention and migration of bone marrow HSCs. Adhesion molecules are involved in both processes regulating hematopoiesis and stem- and progenitor-cell BM retention, migration and development. The mechanisms underlying the movement of stem cells from and to the ... [摘要]  造血干细胞(HSC)由其自我更新的功能能力定义,并分化成所有血细胞谱系。大多数HSC位于骨髓(BM)微环境中的特定解剖位置,处于静止非运动模式。 HSCs与其支持的BM微环境细胞之间的粘附相互作用对于维持干细胞静止和保护免受DNA损伤因子阻止血液学失败和死亡至关重要。多重信号蛋白在控制骨髓HSCs的保留和迁移中起重要作用。粘附分子参与调节造血和干细胞和祖细胞BM保留,迁移和发育的两个过程。干细胞从骨髓移动到骨髓的机制尚未完全阐明,仍然是深入研究的对象。一个重要的方面是修饰干细胞保留,迁移和发育所需的干细胞和祖细胞的粘附分子的表达和亲和力。粘附力通过粘附分子的表达,亲和力和亲合力来调节。亲和力调节与分子结合识别和结合强度有关。在这里,我们描述了在我们的研究中使用的体外 FACS测定来研究整联蛋白α4亚类β1亚单位的表达,亲和力和功能也称为VLA-4)用于小鼠骨髓保留EPCR +长期复制HSC(LT-HSC)(Gur-Cohen等人,2015)。背景整合素是介导细胞和细胞 - ...

Isolation of THY1+ Undifferentiated Spermatogonia from Mouse Postnatal Testes Using Magnetic-activated Cell Sorting (MACS)
Author:
Date:
2016-12-20
[Abstract]  In mammals, homeostasis of many tissues rely on a subpopulation of cells, referred to as stem cells, to sustain an appropriate number of undifferentiated and differentiated cells. Spermatogonial stem cells (SSCs) provide the fundamental cellular source for spermatogenesis and are responsible for the lifelong maintenance of the germline pool in testes throughout the reproductive lifespan of males. To gain insight into germline stem cell biology and develop strategies for infertility treatment, several germ cell isolation methods have been reported in order to acquire good quality and quantity of undifferentiated spermatogonia. Among them, magnetic-activated cell sorting (MACS) is an efficient cell isolation method that requires less time and less initial cell numbers to obtain an enriched ... [摘要]  在哺乳动物中,许多组织的体内平衡依赖于称为干细胞的细胞亚群,以维持适量的未分化细胞和分化细胞。精原细胞干细胞(SSC)为精子发生提供了基本的细胞来源,并且负责终生维持雄性生殖期间睾丸中的种系池。为了获得对种系干细胞生物学的了解并制定不孕症治疗策略,已经报道了几种生殖细胞分离方法,以获得良好质量和数量的未分化精原细胞。其中,磁激活细胞分选(MACS)是一种有效的细胞分离方法,其需要较少的时间和较少的初始细胞数以使用抗原 - 抗体反应来获得富集的细胞群体。胸腺细胞抗原1(THY1,CD90.2)被认为是小鼠新生儿和成年睾丸中未分化精原细胞的表面标志物。在这里,我们描述了一个用于分离来自小鼠睾丸的一周龄THY1 + 细胞和四周龄THY1 + 细胞的方案。分离过程包括三个步骤:睾丸收集和单细胞悬浮,使用生物素缀合的抗THY1抗体的细胞标记和磁性细胞分离。注意,这种隔离方案应在五小时内完成,以最大限度地提高活细胞的质量和数量。

背景 在几个成人组织中观察到活性和静止干细胞的共存。静止,自我更新和分化之间的充分平衡对于维持适当数量的未分化干细胞是必要的,并且避免过早的干细胞耗尽许多组织的体内平衡(Tseng等人,2015; Wabik和Jones,2015; ...

Determination of H2O2 Generation by pHPA Assay
Author:
Date:
2016-11-20
[Abstract]  The production of reactive oxygen species, including H2O2, is a process that can be used in signaling, cell death, or immune response. To quantify oxidative stress in cells, a fluorescence technique has been modified from a previously described method to measure H2O2 release from cells (Panus et al., 1993; Murthy et al., 2010; Larson-Casey et al., 2016; Larson-Casey et al., 2014; He et al., 2011). This assay takes advantage of H2O2-mediated oxidation of horseradish peroxidase (HRP) to Complex I, which, in turn, oxidizes p-hydroxyphenylacetic acid (pHPA) to a stable, fluorescent pHPA dimer (2,2'-dihydroxy-biphenyl-5,5’ diacetate [(pHPA)2]). The H2O2-dependent ... [摘要]  包括H 2 O 2 O 2的活性氧物质的产生是可用于信号传导,细胞死亡或免疫应答的过程。为了量化细胞中的氧化应激,已经从先前描述的方法修改了荧光技术以测量来自细胞的H 2 O 2 O 2释放(Panus等人, 2011; Larson-Casey等人,1993; Murthy等人,2010; Larson-Casey等人,2016; Larson-Casey等人 2014; He 等人,2011)。该测定利用了辣根过氧化物酶(HRP)对配合物I的H 2 O 2 O 2 - 介导的氧化,这又将对羟基苯乙酸(pHPA)氧化成稳定的荧光pHPA二聚体(2,2'-二羟基 - 联苯-5,5'-二乙酸酯[(pHPA)2])。 H 2 O 2 O 2依赖性HRP介导的pHPA氧化是用于定量H 2 O 2 O 2的灵敏和特异性的测定,/sub>从细胞释放。该测定可以测量来自全细胞,线粒体或NADPH氧化酶的H 2 O 2亚型释放。

[背景] (O 2→O 2 - )的超导阴离子(O 2 2-)的歧化导致的强烈的H 2 O 2 O 2 - 非 - 酶促反应,其中所述反应混合物是非酶促的。与O 2 - 不同,H 2 O 2可以容易地穿过膜,因此其能够氧化多个分子。 ...

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