PO.IM01.17 · 免疫学
使用TrailBlazer StarBright Dye抗体偶联试剂盒扩展多重标记能力
Expanding multiplexing capabilities with TrailBlazer StarBright Dye antibody conjugation kits
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
成像、蛋白质印迹和流式细胞术多重标记实验常常受限于某些抗体缺乏合适的荧光基团,从而阻碍了检测组合的规模和最优实验设计。在此,我们使用TrailBlazer™ StarBright™ Dye抗体标记试剂盒来解决这些问题。可使用标记试剂盒生成定制的抗体-染料偶联物,但选择通常局限于标准荧光基团(如FITC),这可能并非最优并限制检测组合的扩展。新推出的TrailBlazer Tag and Label试剂盒能够将抗体偶联至性能更优的StarBright Dyes,提供32种选择,涵盖五条主要激光线。我们的数据显示,由于所采用的新型SpyTag-SpyCatcher技术,使用TrailBlazer试剂盒偶联的抗体可稳定长达一年。这使其非常适合长期储存,并消除了重复标记反应的需要,尤其在纵向研究中可提供可重复的数据。我们还展示了新数据,阐明这些偶联物在各种应用中的多功能性。这包括免疫细胞化学(ICC)染色显微镜数据,通过使用试剂盒偶联的一抗或二抗,它们在多重标记时提供了额外选择。此外,流式细胞术数据显示,检测组合可扩展至纳入荧光基团选择有限的抗体,同时不损害最优检测组合设计,也无需使用二抗。总之,TrailBlazer试剂盒为研究人员带来了重大进展,提供了简便的自偶联方法,并能够在多种应用的复杂生物学研究中生成稳健、可重复的多重标记数据。
查看英文原文 English abstract
Imaging, western blotting, and flow cytometry multiplexing experiments are often constrained by the availability of suitable fluorophores for some antibodies, hindering panel size and optimal experimental design. Here we address those issues using TrailBlazer™ StarBright™ Dye Antibody Labeling Kits. Custom antibody-dye conjugates can be generated using labeling kits, however options are typically restricted to standard fluorophores, such as FITC, which can be suboptimal and limit panel expansion. The newly introduced TrailBlazer Tag and Label Kits have been launched to enable antibody conjugation to superior StarBright Dyes, with 32 options spanning the five main laser lines. Our data shows that antibodies conjugated with TrailBlazer Kits are stable for up to one year, owing to the novel SpyTag-SpyCatcher technology used. This makes them ideal for long-term storage and eliminates the need for repeated labeling reactions, giving reproducible data particularly in longitudinal studies. We also present new data illustrating the versatility of these conjugates in various applications. This includes immunocytochemical (ICC) staining microscopy data, where they provide additional options when multiplexing by using kit-conjugated primary or secondary antibodies. In addition, flow cytometry data show that panels can be expanded to include antibodies that have limited fluorophore options, without compromising optimal panel design and without the use of secondary antibodies. In summary, TrailBlazer Kits offer a significant advance for researchers, delivering an easy self-conjugation method and enabling the generation of robust, reproducible multiplexing data in complex biological studies in multiple applications.
利益披露 Disclosure
S. Sanderson,
Bio-Rad Laboratories Employment.
L. Huang,
Bio-Rad Laboratories Employment.
Y. Kang,
Bio-Rad Laboratories Employment.
R. Mendoza,
Bio-Rad Laboratories Employment.
E. Pusod,
Bio-Rad Laboratories Employment.
M. Blundell,
Bio-Rad Laboratories Employment.
T. Long,
Bio-Rad Laboratories Employment.