| 3-Thiatetradecanoic Acidactivator of PPAR |
Sample solution is provided at 25 µL, 10mM.
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- Purity = 98.00%
- COA (Certificate Of Analysis)
- MSDS (Material Safety Data Sheet)
Chemical structure
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| Cas No. | 116296-31-2 | SDF | Download SDF |
| Synonyms | 3-TDA | ||
| Chemical Name | (undecylthio)-acetic acid | ||
| Canonical SMILES | CCCCCCCCCCCSCC(O)=O | ||
| Formula | C13H26O2S | M.Wt | 246.4 |
| Solubility | ≤30mg/ml in ethanol;30mg/ml in DMSO;30mg/ml in dimethyl formamide | Storage | Store at -20°C |
| Physical Appearance | A crystalline solid | Shipping Condition | Evaluation sample solution : ship with blue ice.All other available size:ship with RT , or blue ice upon request |
| General tips | For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months. | ||
3-Thiatetradecanoic Acid is an activator of PPAR.
The peroxisome proliferator-activated receptors (PPARs) are transcription factors involved in fatty acid metabolism and energy homeostasis. The PPARs also play crucial roles in the control of cellular growth and differentiation.
In vitro: In BT4Cn cells, 3-thiatetradecanoic acid could activate all PPAR subtypes dose-dependently. In cell culture experiments, the PPARγ-selective ligand BRL49653 moderately inhibited growth of BT4Cn cells, while administration of 3-thiatetradecanoic acid led to a marked growth inhibition. Moreover, the administration of the PPARγ-selective antagonist GW9662 abolished BRL49653-induced growth inhibition, but only marginally reduced the effect of 3-thiatetradecanoic acid [1].
In vivo: Administration of 3-thiatetradecanoic acid increased mitochondrial and peroxisomal beta-oxidative capacity and carnitine palmitoyltransferase activity, but reduced free fatty acid and triacylglycerol levels in plasma compared to palmitic acid-treated rats and controls. 3-Thiatetradecanoic acid administration was able to affect the fatty acid composition in plasma and liver by increasing the amount of monoenes [2].
Clinical trial: A previous study described the clinical, hematological, and biochemical safety of 3-thiatetradecanoic acid. 3-Thiatetradecanoic acid was given as a single oral dose for 7 consecutive days. No significant changes were observed in the hematological or clinical chemical parameters in blood/urine. 3-Thiatetradecanoic acid did not induce significant changes in the blood lipids or free fatty acids, but it did lead to an increase in plasma concentration of Δ9 desaturated 3-thiatetradecanoic acid. 3-thiatetradecanoic acid was found to be safe and well tolerated [3].
References:1. Berge K, Tronstad KJ, Flindt EN, Rasmussen TH, Madsen L, Kristiansen K, Berge RK. Tetradecylthioacetic acid inhibits growth of rat glioma cells ex vivo and in vivo via PPAR-dependent and PPAR-independent pathways. Carcinogenesis. 2001 Nov;22(11):1747-55.2. Asiedu, D.K.,Froyland, L.,Vaagenes, H., et al. Long-term effect of tetradecylthioacetic acid: A study on plasma lipid profile and fatty acid composition and oxidation in different rat organs. Biochimica et Biophysica Acta 1300, 86-96 (1996). 3. Pettersen RJ, Salem M, Skorve J, Ulvik RJ, Berge RK, Nordrehaug JE. Pharmacology and safety of tetradecylthioacetic acid (TTA): phase-1 study. J Cardiovasc Pharmacol. 2008 Apr;51(4):410-7.
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逆转录（reverse transcription）是以RNA为模板合成DNA的过程，即RNA指导下的DNA合成。是RNA病毒的复制形式，需逆转录酶的催化。艾滋病病毒(HIV)就是一种典型的逆转录病毒。
逆转录与反转录严格意义上来说没有什么区别，但是逆转录是RNA类病毒自主行为，在整合到宿主细胞内以RNA为模板形成DNA的过程；反转录是进行基因工程过程中，人为地提取出所需要的目的基因的信使RNA，并以之为模板人工合成DNA的过程。二者虽同为RNA→DNA的过程，但地点不同，相对性的来说，逆转录在体内，反转录在体外。
2、取灭过菌且无核酸酶的0.2ml离心管，依次加入2～5μgRNAnμL
3、65℃保温5min，然后冰浴5min；
4、往3步骤中的0.2ml离心管依次加入下列组份
RNase抑制剂（40u/μL）0.5μL
10×M-MLVReactionBuffer2μL
DTT（200mM）1μL
逆转录酶（M-MLV）1μL
5、轻轻混匀后，然后2000rpm离心20s；
6、先在37℃保温1hr，然后70℃保温15min；
7、上述产物可立即进行下一步的PCR反应或-20℃保存。向左转|向右转
在进行RT反应之前，应考虑以下几个方面：
1、RNA
成功的cDNA合成来自高质量的RNA，高质量的RNA至少应保证全长并且不含逆转录酶的抑制剂，如EDTA或SDS。在提取RNA的过程中，要特别防止RNase的污染，同时在逆转录反应中经常加入RNase抑制剂以增加cDNA合成的长度和产量。RNase抑制剂要在第一链cDNA合成反应中，在缓冲液和还原剂（如DTT）存在的条件下加入，因为cDNA合成前的过程会使抑制剂变性，从而释放结合的可以降解RNA的RNase。蛋白RNase抑制剂仅防止RNaseA，B，C对RNA的降解，并不能防止皮肤上的RNase，因此尽管使用了这些抑制剂，也要小心不要从手指上引入RNase，实验过程中经常更换新手套。
2、引物的选择
OligodT
选择OligodT时，要求RNA必须有PolyA，所以真核生物的mRNA都适用。适合长链甚至全长mRNA的RT，所以对RNA样品的质量要求较高，最好不要有明显的DNA污染、RNA降解和RNA断裂。假如想探索新的mRNA进行RT反应，建议推荐使用OligodT引物。使用OligodT引物要比随机引物和特异性引物的稳定性要好。
随机引物
适合各种RNA的RT，尤其适合模板丰度很低的情况（比如某个gene表达量很低）。选择随机引物时，第一链cDNA合成反应中就是以所有的RNA为模板，然后进行PCR反应时设计引物进行特异性扩增。同时要注意随机引物的量和总RNA量之间的关系，一般建议每5μg总RNA的随机引物的用量为50ng，如果每5μg总RNA的随机引物的用量超过250ng，可能会导致小片段产物（<500bp）的增加和长片断、全长产物产物的降低。
特异性引物
特异性引物只能用你设计引物时的下游引物做RT，引物设计质量影响RT的结果，而且不同引物退火温度本来就不相同，所以按照说明书按照一个温度做不是最佳选择，一般不推荐。向左转|向右转
