如何选择合适的流式荧光抗体?
选择错误的抗体可能会浪费时间、金钱和宝贵的样本材料,并可能导致发布不正确或不可复制的数据。选择合适的抗体,研究人员应考虑抗体形式、验证应用和抗原靶标的性质等因素,以增加实验成功的可能性。
在选择用于流式细胞术的抗体时,有许多常见的错误。这些错误包括选择识别错误表位的抗体,例如当目的是避免细胞渗透时,选择靶向细胞内表位的产品;或者选择了错误的克隆,或者选择了不合适的荧光素标记或者不合适的荧光配色方案;研究人员在选择用于流式细胞仪的抗体时还应该考虑的另一个因素是使用直标一抗还是间标方法,目前流式实验直标一抗是首选,此外,直标抗体简化了实验步骤,减少了样品制备过程中发生错误的可能性;目前流式应用主要还是选择单克隆抗体,重组抗体也在渐渐进入大家的选择。这是因为多克隆更容易与靶标以外的蛋白质发生交叉反应,并且经常表现出不同的批次间性能。单克隆抗体,尤其是重组抗体,是一致的,可重复的。一般来说,使用重组产生的抗体可以确保真正的克隆性,减少脱靶结合的机会,并提供尽可能高的批次间再现性。
许多研究人员选择使用与FcγR结合的抗体,从而对所有表达这些受体的细胞进行额外染色。例如,同种型如小鼠IgG2a/b和大鼠IgG2b可通过结合巨噬细胞、单核细胞或b细胞而大大增加背景染色。虽然您可以通过在样品制备中添加Fc阻断剂或血清来解决这个问题,但这可能会导致与其他试剂不兼容。例如,在使用最广泛使用的小鼠CD16/32阻断剂进行治疗后,您无法再对大鼠抗体进行染色,因为阻断剂也是大鼠抗体。小片段F(ab)2抗体没有问题。
有这么多抗体可用,选择合适的产品往往是最重要的。凯靖生物10多年的一级和二级抗体生产经验,专注于抗原抗体的研发和生产,公司构建了基于全自动化高通量杂交瘤和单B细胞技术的单克隆抗体对发现平台,为客户提供创优产品和创新服务。是您值得信赖的现成和定制抗体产品合作伙伴。我们的客户包括许领先的体外诊断制造商、制药公司和研究机构,凭借多年客户服务和经验积累,我们提供始终如一的高质量产品。
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描述 | CD9, also known as Tspan29, is a 24-27 kDa cell surface protein belonging to the tetraspanin family Common to other tetraspanins, CD9 is composed of four transmembrane domains, short N- and C-terminal cytoplasmic domains, and two extracellular loops. The larger extracellular loop, referred to as the LEL or EC2, contains highly conserved CCG and PXSC motifs. The LEL mediates noncovalent protein-protein interactions, allowing tetraspanins to associate with each other as well as signaling molecules, structural proteins, and G-protein coupled receptors. Human CD9 is expressed in multiple cell and tissue types and has been identified in diverse biological roles due to its involvement in the formation of tetraspanin-enriched microdomains (TEMs). TEMs are associated with numerous processes ranging from cell adhesion and fusion, membrane trafficking, and endocytosis to leukocyte adherence and motility. These tetraspanin-enriched microdomains (TEMs) are associated with a wide range of functions from cell adhesion and fusion, membrane trafficking and endocytosis, and eukocyte adherence and motility. The LEL of human CD9 shares 77% and 84% amino acid sequence identity with mouse and rat CD9, respectively. CD9 can form homodimers or interact with other proteins including CD117, CD29, CD46, CD49c, CD81, CD315, Tspan4, TGF-alpha, and HBEGF. Increased expression of CD9 has been shown to enhance transmembrane TGF-alpha -induced EGFR stimulation, and injection of human CD9 mRNA into CD9 knock-out mouse oocytes restored sperm-egg fusion. CD9-LEL may also be involved in the inhibition of multinucleated giant cell formation as well as possess anti-adhesive effects against bacteria trying to invade mammalian cells. CD9 interacts with integrins to regulate cell adhesion and motility. CD9 has been implicated in platelet activation and aggregation. It may act as the terminal signal of myelination in the peripheral nervous system and can regulate the formation of paranodal junctions. Also, it has been suggested CD9 plays an important role both in the self-antigen and recall antigen-induced T cell activation. |
标记 | FITC |
克隆号 | KJ7E6 |
来源宿主 | Recombinant |
克隆类型 | monoclonal |
同种型 | IgG |
特异性 | human |
纯化方式 | Affinity |
激发器(nm) | 488 |
激发光(EX/nm) | 498 |
发射光(Em/nm) | 517 |
存储溶液 | PBS、0.5%BSA、0.03%PC-300 |
存储方式 | Store at 4℃ for one year. |
用法用量 | 5 μl/Test, 0.1 mg/ml |
使用方法 | The procedure is detailed in the document. |