Boc-VPR-AMC，凝血酶和凝血酶-葡萄球菌凝固酶复合物的特异性高荧光底物。凝血酶测定已用于间接测定凝血因子 III。胰蛋白酶、来自大鼠肥大细胞的类胰蛋白酶以及来自海鞘 Halocynthia roretzi 精子的两种胰蛋白酶样酶，acrosin 和精子素，也会水解这种底物。此外，Boc-VPR-AMC 被 Kex2 内切蛋白酶（kexin，以前称为蛋白酶 yscF）切割，这是一种来自酵母 α 细胞的膜结合、钙依赖性丝氨酸蛋白酶。凝血酶的动力学数据：kcat = 105，Km = 21 uM。
编号：165148
CAS号：65147-04-8
单字母：Boc-VPR-AMC
| 编号: | 165148 |
| 中文名称: | 胰凝乳蛋白酶荧光底物BOC-VAL-PRO-ARG-7-氨基-4-甲基香豆素 |
| CAS号: | 65147-04-8 |
| 单字母: | Boc-VPR-AMC |
| 三字母: | Boc N端Boc保护 -Val缬氨酸 -Pro脯氨酸 -Arg精氨酸 -AMC7-氨基-4-甲基香豆素 |
| 氨基酸个数: | 3 |
| 分子式: | C31H45O7N7 |
| 平均分子量: | 627.73 |
| 精确分子量: | 627.34 |
| 等电点(PI): | - |
| pH=7.0时的净电荷数: | 1 |
| 平均亲水性: | 0.75 |
| 疏水性值: | -0.63 |
| 消光系数: | - |
| 来源: | 人工化学合成，仅限科学研究使用，不得用于人体。 |
| 储存条件: | 负80℃至负20℃ |
| 标签: | AMC修饰肽 三肽 血液学 |
Boc-VPR-AMC, a specific, highly fluorogenic substrate for thrombin and the thrombin-staphylocoagulase complex. The thrombin assay has been used for the indirect determination of coagulation factor III. Trypsin, tryptase from rat mast cells, and two trypsin-like enzymes from sperm of the ascidian Halocynthia roretzi, acrosin and spermosin, also hydrolyze this substrate. Furthermore, Boc-VPR-AMC is cleaved by the Kex2 endoprotease (kexin, previously known as proteinase yscF), a membrane-bound, calcium-dependent serine protease from yeast α-cells. Kinetic data for Thrombin: kcat = 105, Km = 21 uM.
多肽荧光标记由于没有放射性，实验操作简单。因此，目前在生物学研究中多肽荧光标记应用非常广泛，多肽荧光标记方法与荧光试剂的结构有关系，对于有游离羧基的采用的方法与接多肽反应相同，也采用HBTU/HOBt/DIEA方法连接。 在N端标记FITC的多肽需经历环化作用来形成荧光素，通常会伴有最后一个氨基酸的去除，但当有一个间隔器如氨基己酸，或者是通过非酸性环境将目的多肽从树脂上切下来时，这种情况可避免在切割的过程中被TFA切割掉。
人们利用利用荧光标记的多肽来检测目标蛋白的活性，并将 其发展的高通量活性筛选方法应用于疾病治疗靶点蛋白的药物筛选和药物开发（例如，各种激 酶、磷酸酶、肽酶等）。
专肽生物能够提供技术成熟的各种荧光标记多肽。
下面是一些常见的多肽修饰荧光物质结构：
FITC标记
FITC(异硫氰酸荧光素)具有比较高的活性，我们公司可以通过两种方式将FITC标记于多肽 上：(1) 将FITC标记于赖氨酸（Lys）或被选择性地脱保护的鸟氨酸（ornithine）侧链氨基 上；(2) 将FITC标记于多肽N端氨基。
当在N端标记时，建议在最后一个氨基和由异硫氰酸酯与氨基反应产生的硫脲键之间引入 烷基间隔器（alkyl spacer），如氨基己酸（Ahx）。链接切割需要酸性环境，在N端标记FITC 的多肽需经历环化作用来形成荧光素，通常会伴有最后一个氨基酸的去除，但当有一个间隔器 如氨基己酸，或者是通过非酸性环境将目的肽从树脂上切下来时，这种情况可避免。空间位阻 被认为是在荧光染料前使用Ahx的主要原因，而不是为什么FITC不能直接偶联在多肽上的原因。
Ahx或b-Ala均可作为间隔器用于FITC标记的多肽上。
| 荧光修饰中文名称 | N端 | N端带有linker |
| 生物素标记多肽 | Biotin- | Biotin-Ahx- |
| 异硫氰酸荧光素 | FITC- | FITC-Ahx- |
| 5-羧基荧光素 | 5-FAM- | 5-FAM-Ahx- |
| 丹磺酰荧光素 | Dansyl- | Dansyl-Ahx- |
| 5-羧基四甲基罗丹明 | TMR- （TAMRA-） | TMR-Ahx- （TAMRA-Ahx-） |
| 多肽N端 | 多肽序列中间 | N端带有linker |
| 生物素标记多肽 | Biotin- | 多肽C端 |
| Lys(Biotin)- | -Lys(Biotin)-- | -Lys(Biotin) |
| Lys(FITC)- | -Lys(FITC)- | -Lys(FITC) |
| Lys(5-FAM)- | -Lys(5-FAM)- | -Lys(5-FAM) |
| Lys(Dansyl)- | -Lys(Dansyl)- | -Lys(Dansyl) |
| Lys(TMR)- | -Lys(TMR)- | -Lys(TMR) |
| Lys(Dnp)- | -Lys(Dnp)- | -Lys(Dnp) |
专肽常做的荧光物质的激发光波长和发射光波长。可供参考选择：
| 荧光基团 | Ex（nm） | Em（nm） | 荧光基团 | Ex（nm） | Em（nm） |
| 羟基香豆素 | 325 | 386 | R-phycoerythrin (PE) (489) | 565 | 578 |
| 丹磺酰氯 | 340 | 578 | Rhodamine Red-X | 560 | 580 |
| AMC | 345 | 445 | Tamara | 565 | 580 |
| 甲氧基香豆素 | 360 | 410 | Alexa fluor 555 | 556 | 573 |
| Alexa fluor 系列 | 345 | 442 | Alexa fluor 546 | 556 | 573 |
| 氨基香豆素 | 350 | 445 | Rox | 575 | 602 |
| Dabcyl | 453 | - | Alexa fluor 568 | 578 | 603 |
| Cy2 | 490 | 510 | Texas Red | 589 | 615 |
| FAM | 495 | 517 | Alexa fluor 594 | 590 | 617 |
| Alexa fluor 488 | 494 | 517 | Alexa fluor | 621 | 639 |
| FITC | 495 | 519 | Alexa fluor 633 | 650 | 668 |
| Alexa fluor 430 | 430 | 545 | Cy5 (625) | 650 | 670 |
| 5-FAM | 492 | 518 | Alexa fluor 660 | 663 | 690 |
| Alexa fluor 532 | 530 | 530 | Cy5.5 | 675 | 694 |
| HEX | 535 | 556 | TruRed | 490; 675 | 695 |
| 5-TAMRA | 542 | 568 | Alexa fluor 680 | 679 | 702 |
| Cy3 | 550 | 570 | Cy7 | 743 | 767 |
| TRITC | 547 | 572 | Cy3.5 | 581 |
596
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