ATP citrate lyase

藤黄提取物和HCA/羟基柠檬酸体外给药抑制HIF激活[2]
使用小鼠视网膜锥细胞系（661W）和人RPE细胞系（ARPE19）通过萤光素酶测定来评估HIF活性，因为光感受器和RPE细胞对AMD的发病机制有显著贡献，尽管AMD患者的iPS细胞衍生的类器官或分化细胞可以被认为是更好的体外系统。在缺氧条件下，HIFα脯氨酰羟化酶（PHD）的活性降低，导致HIFα稳定[12]。加入CoCl2以稳定PHD的抑制作用并激活HIF信号传导。Chetomin被用作HIF抑制剂的阳性对照。我们用藤黄提取物。表1列出了其成分，显示HCA占提取物的一半以上。与对照组相比，藤黄提取物和HCA在ARPE19细胞（图1A）和661W细胞（图1B）中显示出HIF抑制作用。[2]

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藤黄果提取物和HCA/羟基柠檬酸下调Hif1a和下游基因[2]
我们研究了藤黄提取物和HCA如何影响Hif1a和下游基因的mRNA表达。在ARPE19细胞中，无论是否存在CoCl2，施用藤黄提取物都会显著下调Hif1a（图2A）。HIF的下游基因，如Vegfa、Bnip3和Pdk1，被CoCl2上调，被藤黄果提取物给药显著下调（图2B-D）。同样，在661W细胞中施用藤黄提取物后，Hif1a下调（图2E）。在661W细胞中，藤黄提取物的给药也下调了CoCl2诱导的Vegfa上调（图2F）。HIF的其他下游基因的表达与Vegfa一样有下调的趋势（图2G，H）。HCA还下调了ARPE19细胞（图3A-D）和661W细胞（图3E-H）中的Hif1a和下游基因。藤黄提取物和HCA均抑制了ARPE19细胞（图4A，B）和661W细胞（图4C，D）中CoCl2给药后HIF-1α蛋白表达的增加。

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羟基柠檬酸（HCA）是一种经过验证的天然减肥剂，富含藤黄果。（科：Clusiaceae）。本研究使用HPLC和起始密码子靶向（SCoT）分子标记估算的HCA来评估35棵候选加树（CPT）的遗传变异性。还进行了表型和基因型性状之间的关联分析。选定的CPT显示平均HCA含量为29.11 mg/g，Gar 17最高（48.32 mg/g），其次是Gar 6（45.48 mg/g）。SCoT标记分析显示，30个引物中有19个引物共产生151条带，多态性带占66.89%。主坐标分析（PCoA）将CPT组织到散点图的四个象限中，而与HCA含量无关。基于邻域连接方法的树状图通过其自举值证明了其可重复性，并且它有三个聚类。结构分析揭示了所选个体中存在两个假设亚群的可能性。基于一般线性模型（GLM）的关联分析同意SCoT 5d等位基因与HCA含量的强关联，这也支持了Gar 6的前景。基于HCA和SCoT标记的分析有效地追踪了CPT之间的遗传变异和标记-性状关联。据我们所知，这是G.gummi-gutta的第一个发现[5]。

HCA/羟基柠檬酸治疗可以降低肾功能损害的标志物（血尿素氮和血清肌酐）。HCA治疗的小鼠草酸钙晶体沉积明显减少。草酸钙晶体诱导活性氧的产生，降低抗氧化防御酶的活性。HCA减轻了草酸钙结晶引起的氧化应激。HCA对草酸钙诱导的炎性细胞因子如MCP-1、IL-1β和IL-6具有抑制作用。此外，HCA减轻了草酸钙晶体引起的肾小管损伤和细胞凋亡。1.

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给予羟基柠檬酸/HCA抑制了模型小鼠的CNV体积[2]
将悬浮在玉米油中的HCA以30mg/kg/天的剂量腹腔注射共两周，并在开始注射一周后用激光照射小鼠。与对照组相比，HCA给药组在照射第七天的CNV体积显著减少（图6A，B）。

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给予羟基柠檬酸/HCA可抑制体内HIF-1α的表达[2]
将悬浮在玉米油中的HCA腹腔注射（30mg/kg/天）给小鼠共10天，并在给药的第七天用激光照射小鼠。在照射第三天的小鼠视网膜和脉络膜中，HIF-1α因激光照射而增加，因HCA的给药而受到抑制（图7A，B），尽管RPE/脉络膜组织的信号较弱。

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共有135名受试者被随机分配到活性Hydroxycitric acid/羟基柠檬酸组（n=66）或安慰剂组（n=69）；活性羟基柠檬酸组和安慰剂组分别有42例（64%）和42例（61%）完成了12周的治疗（P=0.74）。在12周的治疗期间，两组患者的体重均显著减轻（P<0.001）；然而，组间体重减轻差异没有统计学意义（平均[SD]，3.2[3.3]kg vs 4.1[3.9]kg；P=0.14）。治疗组之间估计的体脂质量损失百分比没有显著差异，受试者体重损失的脂肪比例不受治疗组的影响[4]。

萤光素酶测定[2]
我们使用661W和ARPE19进行了萤光素酶测定，这两种都是转染的HIF萤光素酶报告基因构建体。这些构建体在HRE的控制下编码萤火虫荧光素酶基因，HRE如前所述结合HIF。作为内部对照，这些细胞与CMV肾荧光素酶构建体共转染。我们将0.8×104个细胞/孔/70μL的细胞接种在HTS Transwell®-96接收板上，白色，TC处理，无菌。接种后24小时，200μM CoCl2诱导HIF-αs。将藤黄提取物（藤黄提取物50%（表1））和羟基柠檬酸/HCA溶解在二甲亚砜中，并与CoCl2同时加入生长培养基中。考虑到材料的毒性，我们将溶解在DMSO中的每种化合物加入细胞培养基中，使其浓度为1mg/mL。给药后，细胞在37°C的5%CO2培养箱中孵育24小时。使用Dual luciferase®Reporter检测系统对萤光素酶表达进行定量。荧光强度由微孔板读数器读取。此外，使用100nM的chetomin作为HIF抑制剂的阳性对照，并使用不含CoCl2、藤黄果提取物和HCA的含DMSO培养基作为载体对照。

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蛋白质印迹[2]
在体外实验中，我们向ARPE19细胞系中添加了200μM CoCl2、1 mg/mL藤黄提取物和羟基柠檬酸/HCA，同时考虑了材料的毒性。给药6小时后，将细胞收集在RIPA缓冲液中，并与蛋白酶抑制剂和MG132混合。然后，细胞被均质化。之后，我们对样品进行离心（14800 rpm，4°C，30分钟）并收集上清液。将蛋白质浓度调节至75μg/30μL。

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羟基柠檬酸/HCA的估算[5]
根据Jayaprakasha和Sakariah（2000）的研究，从35个G.gummi gutta CPT的干果皮中提取并纯化HCA（Babu等人，2021；Vishnu等人，2022）。采用高效液相色谱法（HPLC）测定了35个甘米滴胶CPTs中HCA的含量。色谱系统由UFLC（日本京都岛津公司）、SPD-20A检测器、通信总线模块（CBM 20A）和Shim-pack GIST C18柱（5μm，4.6 ID×250 mm）组成。HCA的检测在210 nm处以0.1 AUFS的灵敏度进行。在等度条件下，使用0.6mM硫酸作为流动相，将流速设置为0.7ml/min。从羟基柠檬酸钾中纯化的HCA用作标准品。使用0.45μm PTFE注射器过滤器（I-131 m Axiva；印度Sonipat）过滤标准和样品，并使用20μl注射器将其注入系统。通过绘制不同HCA浓度（200、400、600、800和1000mg/l）与峰面积的关系来制备校准曲线。通过峰积分法估算选定CPT中HCA的浓度，并表示为mg/g样品。

Male C57BL/6J mice were divided into a control group, glyoxylate(GOX) 100 mg/kg group, a GOX+HCA 100 mg/kg group, and a GOX+HCA/Hydroxycitric acid 200 mg/kg group. Blood samples and kidney samples were collected on the eighth day of the experiment. We used Pizzolato staining and a polarized light microscope to examine crystal formation and evaluated oxidative stress via the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the expression of monocyte chemotactic protein-1(MCP-1), nuclear factor-kappa B (NF κB), interleukin-1 β (IL-1 β) and interleukin-6 (IL-6) messenger RNA (mRNA). The expression of osteopontin (OPN) and a cluster of differentiation-44(CD44) were detected by immunohistochemistry and qRT-PCR. In addition, periodic acid Schiff (PAS) staining and TUNEL assay were used to evaluate renal tubular injury and apoptosis. [1]

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Administration of Garcinia Extract and Hydroxycitric acid/HCA to Mice [2]
An MF diet mixed with Garcinia extract at a concentration of 0.2% was administered to 4-week-old male mice for a total of 7 weeks while considering the toxicity of the material. The control group was administered an MF diet. The mice were irradiated with a laser 6 weeks after beginning administration. We injected 30 mg/kg/day HCA suspended in corn oil intraperitoneally to 6-week-old male mice 5 days/week for a total of 2 weeks. The control group was injected with corn oil. The laser was irradiated 1 week after the initial injection.

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Medications in vivo [3]
Hydroxycitric acid tripotassium/K-HCA and citrate acid tripotassium/K-CA were tested as inhibitors to prevent the formation of stones. We randomly divided 600 flies into 6 groups (100 flies in each group) and fed them high-oxalate (0.05% NaOx) medium. Different concentrations (0.01%, 0.1%, and 1%) of K-HCA and K-CA were added in the medium of corresponding groups. Stone formation and life span were assessed in the same way as above.

Adverse Events [1]
No patient was removed from the study protocol for a treatment-related adverse event, and the number of reported adverse events was not significantly different between the placebo and treatment groups (eg, headache, 12 vs 9, respectively; upper respiratory tract symptoms, 13 vs 16, respectively; and gastrointestinal tract symptoms, 6 vs 13, respectively).

[1]. Hydroxycitric acid inhibits renal calcium oxalate deposition by reducing oxidative stress and inflammation. Curr Mol Med. 2020;20(7):527-535.

[2]. Therapeutic Effect of Garcinia cambogia Extract and Hydroxycitric Acid Inhibiting Hypoxia-Inducible Factor in a Murine Model of Age-Related Macular Degeneration. Int J Mol Sci. 2019 Oct 11;20(20). pii: E5049.

[3]. Hydroxycitric Acid Tripotassium Inhibits Calcium Oxalate Crystal Formation in the Drosophila Melanogaster Model of Hyperoxaluria. Med Sci Monit. 2019 May 17;25:3662-3667.

[4]. Garcinia cambogia (hydroxycitric acid) as a potential antiobesity agent: a randomized controlled trial. JAMA. 1998 Nov 11;280(18):1596-600.

[5]. Start codon targeted (SCoT) variability analysis and its association with hydroxy citric acid (HCA) in Garcinia gummi-gutta (L.) Roxb. Plant Gene, Volume 34, June 2023, 100415.

Hydroxycitric acid is a carbonyl compound.
Hydroxycitric acid has been reported in Garcinia cowa, Hibiscus sabdariffa, and Garcinia atroviridis with data available.
See also: Hydroxycitric acid (annotation moved to).

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Background: Age-related macular degeneration (AMD) is the leading cause of blindness and can be classified into two types called atrophic AMD (dry AMD) and neovascular AMD (wet AMD). Dry AMD is characterized by cellular degeneration of the retinal pigment epithelium, choriocapillaris, and photoreceptors. Wet AMD is characterized by the invasion of abnormal vessels from the choroid. Although anti-vascular endothelial growth factor (VEGF) therapy has a potent therapeutic effect against the disease, there is a possibility of chorio-retinal atrophy and adverse systemic events due to long-term robust VEGF antagonism. We focused on hypoxia-inducible factor (HIF) regulation of VEGF transcription, and report the suppressive effects of HIF inhibition against ocular phenotypes in animal models. Many of the known HIF inhibitors are categorized as anti-cancer drugs, and their systemic side effects are cause for concern in clinical use. In this study, we explored food ingredients that have HIF inhibitory effects and verified their effects in an animal model of AMD.
Methods: Food ingredients were screened using a luciferase assay. C57BL6/J mice were administered the Garcinia cambogia extract (Garcinia extract) and Hydroxycitric acid (HCA). Choroidal neovascularization (CNV) was induced by laser irradiation.
Results: Garcinia extract and HCA showed inhibitory effects on HIF in the luciferase assay. The laser CNV model mice showed significant reduction of CNV volume by administering Garcinia extract and HCA. Conclusions: Garcinia extract and HCA showed therapeutic effects in a murine AMD model.
Keywords: Garcinia cambogia; age-related macular degeneration; choroid; hydroxycitric acid; hypoxia-inducible factor; laser induced neovascularization; retina.[2]

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BACKGROUND Hydroxycitric acid is a potential lithontriptic agent for calcium oxalate (CaOx) stones in the kidneys. This study aimed to evaluate the safety and efficiency of hydroxycitric acid tripotassium (K-HCA) against CaOx crystal formation using Drosophila melanogaster hyperoxaluria models. MATERIAL AND METHODS Wild-type D. melanogaster were fed standard medium with ethylene glycol or sodium oxalate added to induce hyperoxaluria. Their Malpighian tubules were dissected and observed under a microscope every 3 days. Crystal deposit score of each Malpighian tubule were evaluated under a magnification of ×200. Using hyperoxaluria Drosophila models, we investigated the inhibitory efficiency of hydroxycitrate acid tripotassium and citric acid tripotassium (K-CA) against CaOx crystal formation. The survival rate of each group was also assessed. RESULTS When fed with 0.05% NaOx, the CaOx formation in Malpighian tubules increased significantly, without reduction of life span. Therefore, we selected 0.05% NaOx-induced hyperoxaluria models for the further investigations. After treatment, the stone scores showed that K-CA and K-HCA both significantly inhibit the formation of CaOx crystals in a dose-dependent manner, and with smaller dosage (0.01%), K-HCA was more efficient than K-CA. Moreover, after treatment of K-CA or K-HCA, the life span in different groups did not change, reflecting the safety to life. CONCLUSIONS The hyperoxaluria Drosophila models fed on 0.05% NaOx diet might be a useful tool to screen novel agents for the management of CaOx stones. K-HCA may be a promising agent for the prevention CaOx stones, with satisfying efficiency and acceptable safety. [3]

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Context: Hydroxycitric acid, the active ingredient in the herbal compound Garcinia cambogia, competitively inhibits the extramitochondrial enzyme adenosine triphosphate-citrate (pro-3S)-lyase. As a citrate cleavage enzyme that may play an essential role in de novo lipogenesis inhibition, G cambogia is claimed to lower body weight and reduce fat mass in humans.
Objective: To evaluate the efficacy of G cambogia for body weight and fat mass loss in overweight human subjects.
Design: Twelve-week randomized, double-blind, placebo-controlled trial.
Setting: Outpatient weight control research unit.
Participants: Overweight men and women subjects (mean body mass index [weight in kilograms divided by the square of height in meters], approximately 32 kg/m2).
Intervention: Subjects were randomized to receive either active herbal compound (1500 mg of hydroxycitric acid per day) or placebo, and both groups were prescribed a high-fiber, low-energy diet. The treatment period was 12 weeks. Body weight was evaluated every other week and fat mass was measured at weeks 0 and 12.
Main outcome measures: Body weight change and fat mass change.
Results: A total of 135 subjects were randomized to either active hydroxycitric acid (n = 66) or placebo (n = 69); 42 (64%) in the active hydroxycitric acid group and 42 (61%) in the placebo group completed 12 weeks of treatment (P = .74). Patients in both groups lost a significant amount of weight during the 12-week treatment period (P<.001); however, between-group weight loss differences were not statistically significant (mean [SD], 3.2 [3.3] kg vs 4.1 [3.9] kg; P = .14). There were no significant differences in estimated percentage of body fat mass loss between treatment groups, and the fraction of subject weight loss as fat was not influenced by treatment group.
Conclusions: Garcinia cambogia failed to produce significant weight loss and fat mass loss beyond that observed with placebo. [4]

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The moderate level of genetic diversity exhibited by the CPTs of G. gummi-gutta supports the precise amplification nature of SCoT markers and the availability of specific SCoT alleles within the species. Interrelationships shown by the individuals irrespective of Hydroxycitric acid/HCA content may be the result of gene flow through migrations. Gar1 was genetically distinct, obviously seen from the scatterplot and dendrogram. STRUCTURE analysis discovered the probability of two assumed subpopulations within the selected individuals. The highest concentration of HCA content was measured in Gar17. However, GLM analysis revealed the promising nature of accession Gar6 as evident by the strong association of its HCA content with SCoT 5d allele. The corresponding allele can significantly contribute to HCA content in the species, which could be taken into consideration while implementing breeding strategies in the species. The findings can be extended for marker-assisted selection and genetic improvement of G. gummi-gutta.[5]

C6H8O8

208.12

208.021

C, 34.63; H, 3.87; O, 61.50

6205-14-7

(-)-Hydroxycitric acid;27750-10-3;Hydroxycitric acid tripotassium hydrate;6100-05-6

123908

Typically exists as solid at room temperature

1.9±0.1 g/cm3

393.3±42.0 °C at 760 mmHg

205.8±24.4 °C

0.0±2.0 mmHg at 25°C

1.620

-1.35

152.36

5

8

5

14

271

0

[K].[K].[K].O([H])C(C(=O)O[H])(C([H])([H])C(=O)O[H])C([H])(C(=O)O[H])O[H] |^1:0,1,2|

ZMJBYMUCKBYSCP-UHFFFAOYSA-N

InChI=1S/C6H8O8/c7-2(8)1-6(14,5(12)13)3(9)4(10)11/h3,9,14H,1H2,(H,7,8)(H,10,11)(H,12,13)

1,2-dihydroxypropane-1,2,3-tricarboxylic acid

hydroxycitric acid; 6205-14-7; Hydroxycitrate; 1,2-dihydroxypropane-1,2,3-tricarboxylic acid; 1,2-Dihydroxy-1,2,3-propanetricarboxylic acid; Pentaric acid, 3-C-carboxy-2-deoxy-; 1,2-dihydroxypropane-1,2,3-tricarboxylicacid; Super CitriMax HCA 600SXS;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。