Suc-Gly-Pro-Leu-Gly-Pro-AMC
目录号 : GA23560
Suc-Gly-Pro-Leu-Gly-Pro-AMC，一种高度敏感的荧光底物，可用于硫氨肽寡肽酶以及脯氨酸后裂解酶（脯氨酰内肽酶）。
Cas No.:72698-36-3
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Experiment[1]: | |
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Sample |
Human plasma |
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Preparation Method |
In each reaction with a total volume of 100 μl of solution, 20 μl of human plasma was added to assay buffer consisting of 20 mM Tris/HCl, pH 8.0, 0.1 M NaCl and 1 mM EDTA. The substrate（Suc-Gly-Pro-Leu-Gly-Pro-AMC） was added to a final concentration of 20 μM. The samples were incubated at 37 °C, and fluorescence was recorded using a microtiter-plate fluorometer with an excitation wavelength of 360 nm and an emission wavelength at 460 nm. |
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Reaction Conditions |
20 μM Suc-Gly-Pro-Leu-Gly-Pro-AMC |
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Applications |
The FAP activity in human plasma was measured using succinyl-pentapeptide composed of SUc-Gly-pro-Leu-Gly-pro-AMC, which contains two FAP cleavage sites.Only the cleavage between the last proline residue and the AMC group releases the fluorescent AMC group, resulting in a fluorescence reading, and the FAP activity of the measured samples ranges from 1.3 to 7 nM/min/ μL. |
| Cell experiment [2]: | |
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Cell lines |
Clone MC3T3-E1 cells |
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Preparation Method |
Samples of 5 x 104 cells were cultured in 35-mm plastic dishes for 5 days. After the cultures were washed with 0.02 M Ca+2- , Mg+2-free phosphate buffer (pH 7.2), they were scraped off from the dishes and homogenized in the same buffer with a glass teflon homogenizer. The homogenate was used for the assay of Suc-Gly-Pro-Leu-Gly-Pro-AMC ase and DAP activities. |
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Reaction Conditions |
30 μl of 2 mM Suc-Gly-Pro-Leu-Gly-Pro-AMC in 37°C for 90 min |
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Applications |
When examining the activities of collagenase-like peptidase ( Suc-Gly-Pro-Leu-Gly-Pro-AMC ase) and dipeptidyl aminopeptidase (DAP) in Mc3t3-e1 in newly cloned osteoblasts with osteogenic ability, the Suc-Gly-Pro-Leu-Gly-Pro-AMC ase in MC3T3-El presented in this paper inhibited by heavy metals (Zn+2 and Cu+2), and some reagents (Mn+2, Ca+2, Mg+2, and EDTA) in creased the enzyme activity; especially, MnanEDTA increased the Suc-Gly-Pro-Leu-Gly-Pro-AMC ase activity to 8.2-fold and 23-fold[3]. |
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References: [1]. Zhen EY, Jin Z, et,al. Circulating FGF21 proteolytic processing mediated by fibroblast activation protein. Biochem J. 2016 Mar 1;473(5):605-14. doi: 10.1042/BJ20151085. Epub 2015 Dec 3. PMID: 26635356; PMCID: PMC4764976. [2]. Chikuma, T., Ishii, Y., Kato, T. et al. Properties of Suc-GPLGP-MCAase and dipeptidyl-aminopeptidase in mouse calvaria-derived osteoblastic cells (MC3T3-E1). Calcif Tissue Int 37, 183–188 (1985). https://doi.org/10.1007/BF02554839 |
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Suc-Gly-Pro-Leu-Gly-Pro-AMC, a highly sensitive, fluorogenic substrate for thimet oligopeptidase as well as for post-proline cleaving enzyme (prolyl endopeptidase) [1].
The FAP activity in human plasma was measured using succinyl-pentapeptide composed of SUch-Gly-pro-Leu-Gly-pro-AMC, which contains two FAP cleavage sites.Only the cleavage between the last proline residue and the AMC group releases the fluorescent AMC group, resulting in a fluorescence reading, and the FAP activity of the measured samples ranges from 1.3 to 7 nM/min/ μL[2].
When each cell suspension was reacted with BODIPY FL casein and seven kinds of peptide-MCA substrates, respectively, a remarkable difference in hydrolytic activities toward Suc-GPLGP-MCA, a substrate toward collagenase-like peptidase, was observed between the constructs: Lc-Triad-displaying cells showed higher catalytic activity than Lc-WT-displaying cells[5].
In Male BALB/c mice ，using Suc-Gly-Pro-Leu-Gly-Pro-AMC test，no significant accumulations of other proteinases, such as matrix metalloproteinases, cathepsin D, and serine proteinases, were determined[3].Suc-GPLGP-MCA is hydro- lyzed at the Leu-Gly bond by CL-peptidase. The CL-peptidase activity in synovial fluid was significantly higher in patients with rheumatoid arthritis (RA) than in patients with osteoarthritis (OA) and in arthropathy-free controls[4].
References:
[1]: Kojima K, Kinoshita H, et,al. A new and highly sensitive fluorescence assay for collagenase-like peptidase activity. Anal Biochem. 1979 Nov 15;100(1):43-50. doi: 10.1016/0003-2697(79)90106-4. PMID: 232383.
[2]: Zhen EY, Jin Z, et,al. Circulating FGF21 proteolytic processing mediated by fibroblast activation protein. Biochem J. 2016 Mar 1;473(5):605-14. doi: 10.1042/BJ20151085. Epub 2015 Dec 3. PMID: 26635356; PMCID: PMC4764976.
[3]: Kakegawa H, Matano Y, et,al. Significant accumulations of cathepsin B and prolylendopeptidase in inflammatory focus of delayed-type hypersensitivity induced by Mycobacterium tuberculosis in mice. Biochem Biophys Res Commun. 2004 Mar 26;316(1):78-84. doi: 10.1016/j.bbrc.2004.01.176. PMID: 15003514.
[4]:Ito A, Hagihara M, et,al. Collagenase-like (CL) peptidase activity in synovial fluid from patients with rheumatoid arthritis. Clin Chim Acta. 1987 Dec;170(2-3):291-6. doi: 10.1016/0009-8981(87)90139-2. PMID: 2830060.
[5]:Okochi N, Kato-Murai M, et,al. Design of a serine protease-like catalytic triad on an antibody light chain displayed on the yeast cell surface. Appl Microbiol Biotechnol. 2007 Dec;77(3):597-603. doi: 10.1007/s00253-007-1197-0. Epub 2007 Sep 27. PMID: 17899065.
Suc-Gly-Pro-Leu-Gly-Pro-AMC，一种高度敏感的荧光底物，可用于硫氨肽寡肽酶和脯氨酸后裂解酶（脯氨酰内肽酶）[1]。
使用由 SUch-Gly-pro-Leu-Gly-pro-AMC 组成的琥珀酰五肽测定人血浆中的 FAP 活性，其中包含两个 FAP 切割位点。仅最后一个脯氨酸残基和 AMC 基团之间的切割释放荧光AMC基团，产生荧光读数，被测样品的FAP活性范围为1.3-7 nM/min/μL[2]。
当每种细胞悬浮液分别与 BODIPY FL 酪蛋白和七种肽-MCA 底物反应时，在构建体之间观察到对 Suc-GPLGP-MCA（胶原酶样肽酶的底物）的水解活性存在显着差异: Lc-Triad展示细胞比Lc-WT展示细胞表现出更高的催化活性[5].
在雄性 BALB/c 小鼠中，使用 Suc-Gly-Pro-Leu-Gly-Pro-AMC 试验，未检测到其他蛋白酶如基质金属蛋白酶、组织蛋白酶 D 和丝氨酸蛋白酶的显着积累 [3].Suc-GPLGP-MCA 在 Leu-Gly 键处被 CL-肽酶水解。类风湿关节炎（RA）患者滑液中的CL-肽酶活性显着高于骨关节炎（OA）患者和无关节病的对照组[4]。
| Cas No. | 72698-36-3 | SDF | |
| 分子式 | C34H44N6O10 | 分子量 | 696.76 |
| 溶解度 | Soluble in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4352 mL | 7.1761 mL | 14.3521 mL |
| 5 mM | 0.287 mL | 1.4352 mL | 2.8704 mL |
| 10 mM | 0.1435 mL | 0.7176 mL | 1.4352 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。