CoraLite® Plus新型荧光染料
照亮您的研究
CoraLite® Plus染料采用超亲水dPEG®技术，是具有极大的亮度和光稳定性的高级荧光团。
CoraLite® Plus新型荧光染料
Proteintech引入的CoraLite® Plus 染料， 采用超亲水dPEG® 基团具有更高的生物相容性和水溶性。该技术也极大提高了染料的亮度和光稳定性。这些染料在光谱上与Alexa Fluor®染料相似。
CoraLite® Plus使低丰度靶点变亮，并且每个珍贵的样品都被计数。CoraLite® Plus 染料偶联抗体可获得更高的信噪比和更低的非特异性结合。这为荧光成像、流式细胞术和蛋白质印迹应用提供了更高的特异性和信号强度。
CoraLite® Plus染料 — 光谱特征
| 荧光染料 |
最大激发波长 (nm) |
最大发射波长 (nm) |
消光系数 (cm-1 M-1) |
光谱相似染料 |
|---|---|---|---|---|
| CoraLite® Plus 405 | 399 | 422 | 35K | Alexa Fluor® 405 |
| CoraLite® Plus 488 | 493 | 522 | 73K | Alexa Fluor® 488, FITC |
| CoraLite® Plus 555 | 554 | 570 | 150K | Alexa Fluor® 555, Cy3, DyLight® 550 |
| CoraLite® Plus 647 | 654 | 674 | 250K | Alexa Fluor® 647, Cy5, DyLight® 650 |
| CoraLite® Plus 750 | 755 | 780 | 240K | Alexa Fluor® 750, Cy7, DyLight® 755 |
什么是dPEG？
聚乙二醇化是将聚乙二醇（PEG）链共价连接到荧光团等各种分子上的过程，以增加其流体动力学体积、水溶性和稳定性。dPEG®（离散PEG）是具有一定数量的环氧乙烷单元的分子。与具有不同大小和分子量的传统PEG相比，它们具有特定的链长、分子量和纯度。不同大小的PEG分子会导致空间位阻和其他问题如结合/相互作用位点问题，从而导致预期活性显著损失。
CoraLite vs CoraLite Plus
Proteintech还提供与传统磺化染料偶联的抗体如CoraLite®。CoraLite®和CoraLite® Plus的主要区别在于CoraLite®染料主链中没有PEG分子。
这两种染料具有相同的激发/发射波长，在光谱上与Alexa Fluor®染料相似。
CoraLite® Plus染料偶联抗体的优势
使用脂肪族或疏水性接头偶联生物分子的方法通常由于接头臂之间的非特异性相互作用而导致最终产物出现问题。超亲水dPEG®技术通过改善特异性相互作用，显著提高最终偶联物的质量。
传统染料标记抗体
- 低标记度（Degree of Labeling，DOL）
- 较高的DOL易导致染料淬灭
- 标记分子在标记或储存过程中易聚集或沉淀
- 易产生非特异性结合
- 背景高
CoraLite® Plus标记抗体
- 高标记度（Degree of Labeling，DOL）
- 染料淬灭少
- 标记分子无聚集或沉淀
- 非特异性结合少
- 信噪比高
dPEG®技术提高了传统染料的偶联效率和亮度
超亲水dPEG® 技术 通过减少负电荷和增加亲水性来提升传统染料。即使相对于传统染料，如FITC、Pacific Blue™和Texas Red®，dPEG®技术也能显著提高标记度和荧光亮度。
FITC vs FITC Plus degree of labeling
- 与传统的FITC染料相比，FITC Plus偶联的抗体标记度更高。
FITC vs FITC Plus brightness
- 与传统的FITC染料相比，FITC Plus偶联的抗体荧光亮度更强。
Proteintech还提供用于流式细胞术的传统染料
- Atlantic Blue™的光谱特性与Pacific Blue™相似。
- Cardinal Red™的光谱特性与Texas Red®相似。
| 荧光染料 |
最大激发波长 (nm) |
最大发射波长 (nm) |
消光系数 (cm-1 M-1) |
光谱相似染料 |
|---|---|---|---|---|
| Atlantic Blue™ | 404 | 458 | 19K | Pacific Blue™ |
| FITC/FITC Plus | 495 | 524 | 76K | Alexa Fluor® 488 |
| PE | 566 | 574 | 1,960K | Alexa Fluor® 555, Cy3, DyLight® 550 |
| Cardinal Red™ | 592 | 611 | 80K | Alexa Fluor® 594, Texas Red® |
| APC | 650 | 661 | 700K | Alexa Fluor® 647, Cy5, DyLight® 650 |
CoraLite® Plus偶联抗体在各种应用中表现出卓越的性能
Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using CoraLite® Plus 488 MAP2 antibody (CL488-17490) at dilution of 1:200, CoraLite 594 GFAP antibody (CL594-16825, red). DAPI (blue).
Immunofluorescent analysis of (4% PFA) fixed sodium arsenite treated HeLa cells using CoraLite® Plus 488 G3BP1 antibody (CL488-13057) at dilution of 1:200, CL594-Phalloidin (red).
Immunofluorescence of human kidney: FFPE human kidney sections were stained with anti-Calbindin antibody (14479-1-AP) labeled with FlexAble CoraLite® Plus 555 Kit (KFA002, yellow), anti-ACE2 antibody (66699-1-Ig) labeled with FlexAble CoraLite® Plus 647 Kit (KFA023, magenta), CoraLite® Plus 488-conjugated Podocalyxin antibody (CL488-18150, green), and DAPI (blue).
WB of HeLa cell lysates: HeLa cell lysates were detected with anti-GAPDH antibody (10494-1-AP) labeled with FlexAble CoraLite® Plus 555 Kit (KFA002, green) and anti-NUDT21 antibody (66335-1-Ig) labeled with FlexAble CoraLite® Plus 647 Kit (KFA023, red).
Various lysates were subjected to SDS PAGE followed by western blot with CL594-66009 ( Beta Actin antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
Various lysates were subjected to SDS PAGE followed by western blot with CL488-66528 ( IDO1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
Flow cytometry of PBMC. 1X10^6 human peripheral blood mononuclear cells (PBMCs) were stained with 0.5 µg anti-human CD3 antibody (clone UCHT1, 65151-1-Ig) labeled with FlexAble CoraLite® Plus 405 Kit (KFA026).
1X10^6 human PBMCs were surface co-stained with FITC Anti-Human CD45RO and 5 ul CoraLite® Plus 405 Anti-Human CD45RA (CL405-65226, Clone:F8-11-13) or Mouse IgG1 Isotype Control. Cells were not fixed. Lymphocytes were gated.
1X10^6 human PBMCs were surface co-stained with PE Anti-Human CD3 and 5 ul CoraLite® Plus 750 Anti-Human CD4 (CL750-65134, Clone:OKT4) or Mouse IgG2b Isotype Control. Cells were not fixed. Lymphocytes were gated.
相关内容
Alexa Fluor® is a registered trademark of Life Technologies Corporation.
DyLight® is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
Pacific Blue™ is a trademark of Molecular Probes, Inc.
dPEG® is a trademark from Quanta Biodesign Ltd
