Natural flavonoid from green tea

(+)-Gallocatechin（10-300 μM，48 小时）会导致细胞凋亡并阻止 HCT-116 和 SW-480 细胞增殖 [1]。

细胞凋亡分析 [1]
细胞类型： HCT-116 细胞
测试浓度： 0-100 µM
孵育时间： 48 h
实验结果： 浓度为 60、80 和 100 µM 时的细胞凋亡率：14.1%、25.0%、23.5%。

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细胞增殖分析[1]
茶多酚溶解在DMSO中，使用前储存在-20°C。将细胞接种在96孔板中（1×10~4个细胞/孔）。24小时后，向孔中加入指定浓度的药物。DMSO的终浓度为1%。对照组暴露于含有1%DMSO的培养基中，不含药物。在指定的潜伏期后，根据制造商的说明，使用MTS测定法评估细胞增殖。简而言之，在每个孔中用100μL新鲜培养基和20μL MTS试剂（CellTiter 96水溶液）替换培养基，并将平板放回培养箱1-2小时。将每个孔中的60μL等分培养基转移到ELISA 96孔平板上，记录其在490 nm处的吸光度。由于1%DMSO不影响两种细胞系的增殖，因此结果以对照的百分比表示（DMSO载体设置为100%）。

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细胞周期分析[1]
将HCT-116细胞以2×10~5个细胞的密度铺在24孔组织培养板上。培养1天后，更换培养基并加入茶多酚。细胞在收获前孵育48小时。将这些细胞在冰箱中用80%乙醇轻轻固定2小时，然后在冰浴中用0.25%Triton X-100处理5分钟。将细胞重新悬浮在150µL含有40µg/mL碘化丙啶（PI）和0.1 mg/mL RNase的PBS中。将细胞在室温下在暗室中孵育20分钟，并使用FACScan流式细胞仪 和FlowJo 9.0软件进行细胞周期分析。对于每次测量，至少计数10000个细胞。

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细胞凋亡分析[1]
将HCT-116细胞接种在24孔组织培养板上（2×10~5个细胞/孔）。第二天，更换培养基，用试验化合物处理细胞。处理48小时后，收集悬浮在培养基中的细胞。用0.05%胰蛋白酶分离贴壁细胞。然后加入含有FBS和漂浮细胞的培养基以灭活胰蛋白酶。轻轻移液后，将细胞在1500×g下离心5分钟。去除上清液，根据制造商的说明用膜联蛋白V-FITC和PI对细胞进行染色。膜联蛋白V-FITC在细胞凋亡早期检测磷脂酰肌醇从细胞内膜向外膜的移位，PI可以在细胞凋亡晚期或坏死时进入细胞。使用未处理的细胞作为双重染色的对照。染色后立即使用FACScan流式细胞仪和FlowJo 9.0软件分析细胞。对于每次测量，至少计数20000个细胞。

[1]. Epigallocatechin Gallate (EGCG) is the most effective cancer chemopreventive polyphenol in green tea. Nutrients. 2012 Nov 8;4(11):1679-91.

Gallocatechin is a catechin that is a flavan substituted by hydroxy groups at positions 3, 3', 4', 5, 5' and 7 (the trans isomer). It is isolated from Acacia mearnsii. It has a role as a metabolite. It is a catechin and a flavan-3,3',4',5,5',7-hexol.
(+)-Gallocatechin has been reported in Camellia sinensis, Quercus acutissima, and other organisms with data available.
See also: Cianidanol (subclass of); Crofelemer (monomer of); Green tea leaf (part of).

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Green tea is a popular drink consumed daily by millions of people around the world. Previous studies have shown that some polyphenol compounds from green tea possess anticancer activities. However, systemic evaluation was limited. In this study, we determined the cancer chemopreventive potentials of 10 representative polyphenols (caffeic acid, CA; gallic acid, GA; catechin, C; epicatechin, EC; gallocatechin, GC; catechin gallate, CG; gallocatechin gallate, GCG; epicatechin gallate, ECG; epigallocatechin, EGC; and epigallocatechin gallate, EGCG), and explored their structure-activity relationship. The effect of the 10 polyphenol compounds on the proliferation of HCT-116 and SW-480 human colorectal cancer cells was evaluated using an MTS assay. Cell cycle distribution and apoptotic effects were analyzed by flow cytometry after staining with propidium iodide (PI)/RNase or annexin V/PI. Among the 10 polyphenols, EGCG showed the most potent antiproliferative effects, and significantly induced cell cycle arrest in the G1 phase and cell apoptosis. When the relationship between chemical structure and anticancer activity was examined, C and EC did not show antiproliferative effects, and GA showed some antiproliferative effects. When C and EC esterified with GA to produce CG and ECG, the antiproliferative effects were increased significantly. A similar relationship was found between EGC and EGCG. The gallic acid group significantly enhanced catechin's anticancer potential. This property could be utilized in future semi-synthesis of flavonoid derivatives to develop novel anticancer agents.[1]

C15H14O7

306.2675

306.073

C, 58.83; H, 4.61; O, 36.57

970-73-0

(-)-Epigallocatechin;970-74-1;(+)-Gallocatechin-13C3

65084

White to off-white solid powder

1.7±0.1 g/cm3

685.6±55.0 °C at 760 mmHg

189 - 191 °C

368.5±31.5 °C

0.0±2.2 mmHg at 25°C

1.776

-0.1

130.61

6

7

1

22

380

2

C1[C@@H]([C@H](OC2=CC(=CC(=C21)O)O)C3=CC(=C(C(=C3)O)O)O)O

XMOCLSLCDHWDHP-SWLSCSKDSA-N

InChI=1S/C15H14O7/c16-7-3-9(17)8-5-12(20)15(22-13(8)4-7)6-1-10(18)14(21)11(19)2-6/h1-4,12,15-21H,5H2/t12-,15+/m0/s1

(2R,3S)-2-(3,4,5-trihydroxyphenyl)-3,4-dihydro-2H-chromene-3,5,7-triol

(+)-Gallocatechin; Gallocatechin; 970-73-0; Gallocatechol; (+)-gallocatechol; d-Gallocatechin; 1617-55-6; (2R,3S)-gallocatechin;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~326.51 mM)
H2O : ~3.33 mg/mL (~10.87 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (8.16 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (8.16 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (8.16 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。