
Product information | Key steps | References | Technical support | Reviews |
Product overview
Product name | Kynurenic Acid/Quinolinic Acid ELISA pack |
Description | Two enzyme immunoassays (ELISA) allowing the quantitative determination of Kynurenic acid and Quinolinic acid in serum samples. |
Labels | CE |
Format | 2 x 96-well plate |
Samples | Serum, Plasma |
Minimum sample volume | 25-50 µL |
Cross specie-reactivity | React with all species |
Standard range | KYNA: 1.40 - 74 ng/ml QA: 25 - 2000 ng/ml |
Sensitivity | LoD KYNA: 0.53 ng/ml LoD QA: 6 ng/ml |
Specificity | No significant cross-reactivity was observed analogs for each kit. See product pages for Kynurenic acid ELISAand Quinolinic acid ELISA. |
Storage | Store at 2-8°C for to 6 months. |
Datasheets | Instructions for use - KYNA kit, Instructions for use - QA kit Safety datasheet - KYNA kit, Safety datasheet QA kit |
Key steps
Sample collection & storage | Serum: Do not use lipemic, haemolytic samples, as well as samples containing precipitates or fibrin strands. Store samples at 2-8°C for up to 48h or -20°C for longer period (up to 6 months). |
Sample preparation | KYNA: Sample derivatization (90 min) QA: Sample acylation (120 min) |
ELISA | Antisera overnight incubation, revelation and read steps (1h). |
Detailed protocol | Instructions for use - KYNA kit, Instructions for use - QA kit |
References
Selected articles on Kynurenic acid:
- Pedraz-Petrozzi et al. Effects of inflammation on the kynurenine pathway in schizophrenia - a systematic review. J Neuroinflammation. 2020
- Achtyes et al. Inflammation and kynurenine pathway dysregulation in post-partum women with severe and suicidal depression. Brain Behav Immun. 2020
- Walczak et al. Kynurenic acid and cancer: facts and controversies. Cell Mol Life Sci. 2019
- Lim et al. Kynurenine pathway metabolomics predicts and provides mechanistic insight into multiple sclerosis progression. Sci Rep. 2017
- Plitman et al. Kynurenic Acid in Schizophrenia: A Systematic Review and Meta-analysis. Schizophr Bull. 2017
- Wirthgen et al. Kynurenic Acid: The Janus-Faced Role of an Immunomodulatory Tryptophan Metabolite and Its Link to Pathological Conditions. Front Immunol. 2017
Selected articles on Quinolinic acid:
- Fertan et al. Effects of the Novel IDO Inhibitor DWG-1036 on the Behavior of Male and Female 3xTg-AD Mice. Front Pharmacol. 2019 Sep;24
- Pierozan et al. Synergistic Toxicity of the Neurometabolites Quinolinic Acid and Homocysteine in Cortical Neurons and Astrocytes: Implications in Alzheimer"s Disease. Neurotox Res. 2018 Jul;34
- Hernandez-Martinez et al. Quinolinic acid induces neuritogenesis in SH-SY5Y neuroblastoma cells independently of NMDA receptor activation. Eur J Neurosci. 2017 Mar;45
- Sundaram et al. Quinolinic acid toxicity on oligodendroglial cells: relevance for multiple sclerosis and therapeutic strategies. J Neuroinflammation. 2014 Dec 13
- Guillemin GJ. Quinolinic acid, the inescapable neurotoxin. FEBS J. 2012 Apr;279
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氯仿萃取RNA
异丙醇沉淀RNA
酒精清洗
--本来,看好丹麦Exiqon公司,Qiagen的两个试剂盒,无奈经费有限,这两个盒子很不错,价格也不菲,有米的兄弟姐妹可以买。
--另外查询到国产上海诺伦公司的血液(血清/血浆)总RNA抽提试剂盒--广州这边用的人不多,不知道上海那边的兄弟姐妹有什么经验?
---还有,北京康为世纪公司的游离RNA(血清血浆尿液)提取试剂,货号:CW2281,也比较便宜,但是俺心中也没有底,请大家给点意见。
---已经订购了LIFETECHNOLOGY的TrizolLS专门提取体液RNA的试剂(100毫升2000元人民币,也不菲),准备预试验看看提取RNA的质量。(Lifetechnology公司的AM1556也不错,但是也不便宜,每个盒子40次/2590元)
----但是还是希望有试剂盒,比较简单,快捷,最重要的是样本量太大,要分离400份左右的血清。
TRIZOL的主要成分是苯酚。苯酚的主要作用是裂解细胞,使细胞中的蛋白,核酸物质解聚得到释放。苯酚虽可有效地变性蛋白质,但不能完全抑制RNA酶活性,因此TRIzol中还加入了8-羟基喹啉、异硫氰酸胍、β-巯基乙醇等来抑制内源和外源RNase(RNA酶)。
TRIZOL是从细胞和组织中提取总RNA的即用型试剂,在样品裂解或匀浆过程中,TRIZOL 能保持RNA完整性。加入氯仿后,溶液分为水相和有机相,RNA在水相中。取出水相,用异丙醇可沉淀回收RNA。
※0.1%的8-羟基喹啉可以抑制RNase,与氯仿联合使用可增强抑制作用。
※异硫氰酸胍属于解偶剂,是一类强力的蛋白质变性剂,可溶解蛋白质并使蛋白质二级结构消失,导致细胞结构降解,核蛋白迅速与核酸分离。
※β-巯基乙醇的主要作用是破坏RNase蛋白质中的二硫键。
TRIZOL是一种新型总RNA抽提试剂,可以直接从细胞或组织中提取总RNA。其含有苯酚、异硫氰酸胍等物质,能迅速破碎细胞并抑制细胞释放出的核酸酶。TRIZOL在破碎和溶解细胞时能保持RNA的完整性,因此对纯化RNA及标准化RNA的生产十分有用。
基本特点
Trizol试剂可以快速提取人、动物、植物、细菌不同组织的总RNA,该方法对少量的组织(50-100 mg)和细胞(5×106)以及大量的组织(≥1 g)和细胞(>107)均有较好的分离效果。TRIZOL试剂操作上的简单性允许同时处理多个的样品。所有的操作可以在一小时内完成。TRIZOL抽提的总RNA能够避免DNA和蛋白的污染。故而能够作RNA 印迹分析、斑点杂交、poly(A)+ 选择、体外翻译、RNA酶保护分析和分子克隆。并且利用DNA、RNA和蛋白质在不同溶液中的溶解性质,可以通过分层分别将不同层中的RNA(上层)、DNA(中层)、蛋白质(下层)分离纯化出来,效率极好。
Trizol试剂能促进不同种属不同分子量大小的多种RNA的析出。例如,从大鼠肝脏抽提的RNA琼脂糖凝胶电泳并用溴化乙啶染色,可见许多介于7 kb和15 kb之间不连续的高分子量条带,(mRNA和hnRNA成分)两条优势核糖体RNA条带位于~5 kb (28S)和~2 kb (18S),低分子量RNA介于0.1 和 0.3 kb之间 (tRNA,5S)。当抽提的RNA用TE稀释时其A260/A280比值≥1.8
TRIZOL
http://baike.baidu.com/view/533842.htm
Trizol法和试剂盒提取组织中的RNA,哪个方法更可靠和精确呢?谢谢!
这里有关于miRNA专题报道,可以看看
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