Fluorogenic Substrates
haemtech含氟底物
基于ANSN的成花底物的一般結(jié)構(gòu)
闡述了6-氨基-1-萘磺酰胺（ANSN）熒光底物的一般結(jié)構(gòu)。在目前的化合物家族中，R1是一種三肽，其COOH末端殘基通常是精氨酸。R2和R3可以是氫、烷基、芳基或環(huán)烷基。虛線表示各種絲氨酸蛋白酶水解的位置。
含有熒光報(bào)告基團(tuán)6-氨基-1-萘磺酰胺（ANSN）的底物是監(jiān)測(cè)各種絲氨酸蛋白酶（1-12）酶活性的有用化合物。在這類化合物中，ANSN報(bào)告基團(tuán)（在R1位置）與短的三肽序列相連。肽序列的設(shè)計(jì)是為了優(yōu)化酶和底物之間的相互作用?？梢蕴砑拥絉2和R3位置的附加組分反映了P'亞基位置的變化，并且通常會(huì)影響底物的動(dòng)力學(xué)參數(shù)。作為有效底物的化合物在三肽和ANSN基團(tuán)之間水解。一旦從肽段分離，ANSN基團(tuán)的相對(duì)熒光強(qiáng)度增加了約1000倍。
下頁(yè)確定的動(dòng)力學(xué)特性將有助于選擇合適的底物。ANSN底物被證明特別適用于因子VIIa（1-5,7,9,10,12）的分析。盡管僅因子VIIa的底物水解速率相對(duì)較慢，但當(dāng)將組織因子納入分析系統(tǒng)時(shí)，化合物SN-17a等幾種化合物的kcat發(fā)生了很大變化。
使用信息：
在二甲基亞砜中，以10mM的儲(chǔ)備溶液形式提供基于ANSN的底物，通常用于分析濃度范圍為400 nM至50pM的酶。ANSN底物通常在50至100µM的終濃度下使用，并且可以在大多數(shù)生理緩沖液（即TBS、HBS、PBS等）中稀釋到該終濃度。反應(yīng)（熒光的相對(duì)變化）可使用標(biāo)準(zhǔn)熒光計(jì)或熒光板閱讀器進(jìn)行監(jiān)測(cè)。儀器設(shè)置在以下波長(zhǎng)（激發(fā)=352nm，發(fā)射=470nm）時(shí)，可以使用動(dòng)力學(xué)或端點(diǎn)模式。通過(guò)在發(fā)射光束中使用390到450nm長(zhǎng)通截止濾波器，可以將光偽影小化。
下表列出了每種基質(zhì)的動(dòng)力學(xué)特性，下表列出了使用實(shí)例的期刊參考文獻(xiàn)列表。
二甲基亞砜中的儲(chǔ)備基質(zhì)溶液應(yīng)在4°C或更低溫度下冷凍保存，并應(yīng)避光。在這些條件下，化合物將保持穩(wěn)定一年以上。
**下表關(guān)鍵字：Km=µM；kcat=s-1；kcat/Km=M-1s-1 x 10-5；ND=未確定
- SN-5 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | D-AFK-ANSNH-iC4H9.2HBr |
|---|
| Useful For | Useful for plasmin |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
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| KM | ND | ND | ND | ND | ND | ND | ND | 130 |
| Kcat | ND | ND | ND | ND | ND | ND | ND | 3.7 |
| Kcat/KM | ND | ND | ND | ND | ND | ND | ND | 0.28 |
- SN-7 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | Mes-D-LGR-ANSN(C2H5)2 |
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| Useful For | Useful for factor Xa |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
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| KM | 31 | 180 | 200 | 125 | 580 | 113 | ND | ND |
| Kcat | 0.63 | 0.007 | 0.79 | 36 | 15 | 0.055 | ND | ND |
| Kcat/KM | 0.2 | ND | 0.04 | 2.9 | 0.26 | 0.005 | ND | ND |
- SN-13a 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | D-LPR-ANSNH-C3H7 • 2 HCl |
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| Useful For | Useful for factor XIa |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
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| KM | 0.5 | 300 | 300 | 171 | 75 | 45 | 98 | ND |
| Kcat | 19 | 0.07 | 4.5 | 3.3 | 53 | 52 | 0.31 | ND |
| Kcat/KM | 380 | 0.002 | 0.15 | 0.19 | 7.1 | 12 | 0.03 | ND |
- SN-17a 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | D-FPR-ANSNH-C6H11 • 2 HCl |
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| Useful For | Useful for Thrombin & VIIa |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
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| KM | 0.4 | 150 | 330 | 150 | ND | 7.8 | 36 | ND |
| Kcat | 17 | 0.05 | 8.4 | 0.32 | ND | 6.6 | 0.074 | ND |
| Kcat/KM | 430 | 0.004 | 0.25 | 0.02 | ND | 8.6 | 0.02 | ND |
- SN-17c 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | D-FPR-ANSNH-C4H9 • 2 HCl |
|---|
| Useful For | Useful for factor VIIa |
|---|
| Formulation | DMSO |
|---|
| Concentration | 10 mM |
|---|
| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
|---|
| KM | ND | 186 | 102 | ND | ND | 53 | ND | ND |
| Kcat | ND | 0.11 | 2.7 | ND | ND | 21 | ND | ND |
| Kcat/KM | ND | 0.006 | 0.26 | ND | ND | 4 | ND | ND |
- SN-18 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
SAFETY DATA SHEET
| Structure | Boc-L-(p-F)FPR-ANSNH-C2H5 |
|---|
| Useful For | Useful for tPA |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
|---|
| KM | 3.7 | 50 | 217 | ND | ND | ND | 71 | ND |
| Kcat | 44 | 0.008 | 0.88 | ND | ND | ND | 1.03 | ND |
| Kcat/KM | 120 | 0.002 | 0.04 | ND | ND | ND | 0.15 | ND |
- SN-20 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
SAFETY DATA SHEET
| Structure | Boc-L-FPR-ANSNH-C2H5 |
|---|
| Useful For | Useful for thrombin |
|---|
| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
|---|
| KM | 17 | ND | ND | 100 | ND | 540 | 47 | ND |
| Kcat | 58 | ND | ND | 0.31 | ND | 2.2 | 0.011 | ND |
| Kcat/KM | 34 | ND | ND | 0.003 | ND | 0.04 | 0.002 | ND |
- SN-45 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
SAFETY DATA SHEET
| Structure | L-EGR-ANSNH-C3H7 • 2 HBr |
|---|
| Useful For | Useful for factor XIa |
|---|
| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
|---|
| KM | 100 | ND | ND | 110 | 225 | 440 | ND | ND |
| Kcat | 2.5 | ND | ND | 0.2 | 82 | 17 | ND | ND |
| Kcat/KM | 0.25 | ND | ND | 0.02 | 3.6 | 0.39 | ND | ND |
- SN-54 6-ami-naphthalenesulfonamide-based (ANSN) fluorogenic substratePricing: Please Inquire
| Structure | BOC-D-VLR-ANSNH-C4H9 |
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| Useful For | Useful for aPC |
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| Formulation | DMSO |
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| Concentration | 10 mM |
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| Kinetic Properties | IIa | VIIa | VIIa/TF | Xa | XIa | aPC | tPA | Plasmin |
|---|
| KM | 19 | 42 | 170 | 19 | ND | 3.9 | ND | ND |
| Kcat | 0.055 | 0.007 | 1.6 | 0.055 | ND | 2.1 | ND | ND |
| Kcat/KM | 0.03 | 0.002 | 0.09 | 0.03 | ND | 5.2 | ND | ND |
