- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Key Benefits
- Non Radioactive.
- Can monitor multiple time points to follow kinetics.
- One-step, no wash assay.
- Adaptable for High Throughput format.
- Sensitive.
- Applications – Fluorescent Plate Reader.
Additional information
| Kit Size | 500 |
|---|
Monoamine oxidase (MAO) is a flavin-containing enzyme that catalyses the oxidation of a variety of amine-containing neurotransmitters such as serotonin, norepinephrine, epinephrine and dopamine to yield the corresponding aldehydes (1). MAO exists in two isoforms, namely MAO-A and MAO-B, which are the products of two distinct genes (2). MAO-A and B exhibit different specificities to substrates and inhibitor selectivities. Extensive studies have been preformed to characterize their properties (3-7). MAO-A acts preferentially on serotonin and norepinephrine, and is inhibited by clorgyline. MAO-B acts preferentially on 2-phenylethylamine and benzylamine and is inhibited by deprenyl and pargyline. Localized in the outer mitochondrial membrane, these enzymes are found throughout the body. Often only one form of the enzyme is present in a specific organ and/or within a specific cell type (8-9). In addition to their role in regulating neurotransmitters, these enzymes are also involved in processing biogenic amines (10) including tyramine (11). The Fluoro MAO-A/B detection kit utilizes a non – fluorescent substrate, 10-Acetyl-3, 7-dihydroxyphenoxazine (ADHP) to detect H202 released from the conversion of a substrate to its aldehyde via MAO-A/B. Furthermore, H202 oxidizes ADHP in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase.
| Document Title |
| Fluoro MAO Protocol |
| Fluoro MAO Datasheet |
| msds.fluoroMAO |
| Reference |
| Waldmeier PC (1987) Amine oxidases and their endogenous substrates. J Neural Transm Suppl 23:55–72. |
| Bach, A. W. J., N. C. Lan, D. L. Johnson, C. W. Abell, M. E. Bembenck, S. W. Kwan, P. H. Seeburg & J. C. Shih: cDNA clon-ing of human liver monoamine oxidase A and B: molecular basis of differences in enzymatic properties. Proc. Nat. Acad. Sci. U.S.A. 1988, 85, 4934–4938. |
| Johnston, J. P.: Some observations upon a new inhibitor of mono-amine oxidase in brain tissue. Biochem. Pharmacol. 1968, 17, 1285–1297. |
| Suzuki, O., E. Noguchi & K. Yagi: A simple fluorometric assay for type B monoamine oxidase activity in rat tissues. J. Biochem. 1976, 79, 1297–1299. |
| Fowler, C. J. & B. A. Callingham: Substrate-selective activation of rat liver mitochondrial monoamine oxidase by oxygen. Biochem. Pharmacol. 1978, 27, 1995–2000. |
| Tipton, K. F.: Enzymology of monoamine oxidase. Cell Biochem. Funct. 1986, 4, 79–87. |
| Youdim, M. B. H. & M. Tenne: Assay and purification of liver monoamine oxidase. Methods Enzymol. 1987, 142, 617–626. |
| Trendelenburg U, Cassis L, Grohmann M and Langeloh A (1987) The functional coupling of neuronal and extraneuronal transport with intracellular monoamine oxidase. J Neural Transm Suppl 23:91–101. |
| Yu PH, Davis BA and Boulton AA (1992) Neuronal and astroglial monoamine oxidase: Pharmacological implications of specific MAO-B inhibitors. Prog Brain Res 94:309–315. |
| Strolin Benedetti M and Tipton KF (1998) Monoamine oxidases and related amine oxidases as phase I enzymes in the metabolism of xenobiotics. J Neural Transm Suppl 52:149–171. |
| Hauptmann N, Grimsby J, Shih JC and Cadenas E (1996) The metabolism of tyramine by monoamine oxidase A/B causes oxidative damage to mitochondrial DNA. Arch Biochem Biophys 335:295–304. Methods Enzymology: Metabolism of Aromatic Amino Acids and Amines. Volume 142, page 617 (1997).13. Holt A. ; Sharman D.F. ; Baker G.B. ; Palcic M.M. A Continuous Spectrophotometric Assay for Monoamine Oxidase and Related Enzymes in Tissue Homogenates Analytical Biochemistry, January 1997, vol. 244, no. 2, pp. 384-392(9). |
| D. W. R. Hall, Bridget W. Logan and G. H. Parsons. Further studies on the inhibition of monoamine oxidase by M & B 9302 (clorgyline)—I .Substrate specificity in various mammalian species. Biochemical Pharma |
| Part# | Reagent | Temperature |
| Part # 3020 | 5X Reaction Buffer, 1 Bottle | 2-8C |
| Part # 6007 | Horseradish Peroxidase, 1 Vial | 2-8C |
| Part # 7006 | MAO-B Substrate, Benzylamine, 1 Vial | 2-8C (Aliquot in Single Use Vials) |
| Part # 7005 | MAO-A/B Substrate, Tyramine, 1 Vial | 2-8C (Aliquot in Single Use Vials) |
| Part # 7002 | Clorgyline: Monoamine Oxidase A Inhibitor, 1 Vial | 2-8C (Aliquot in Single Use Vials) |
| Part # 7003 | Pargyline: Monoamine Oxidase B Inhibitor, 1 Vial | -20C (Aliquot in Single Use Vials) |
| Part# 4009 | Detection Reagent, 1 Vial | -20C (Aliquot in Single Use Vials) |
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利用NheI和HindIII双酶切插入目的片段,为什么双酶切有目的片段和切开的质粒,测序却没有?
并且测序后在NheI位点后的序列变成了-GGCTAGGTAC-Kpn位点(CGTTTAAACTTAAGCTTG消失。之前和之后的序列都相符),之间的怎么都没了?
请教高人,这样的问题怎么解决?
多谢
你所说的“测序酶”在一般情况下就是“聚合酶”。
因为目前的的测序方法主要就是借助聚合反应。
当然,有些测序方法(比如曾经的SOLiD系统)是利用连接反应,那么它用的“测序酶”肯定是连接酶。
估计,也就是这个原因有人把它。
两个基因大小分别230kb和450kb,酶切后,450kb的有目的条带,但很弱,230kb的质粒条带亮,其下方有一很微弱条带,用的酶分别是:Ncol和Spel体系是(Takara,两个酶切体系不同,用的官网推荐体系):
NcoI1μl
Spel1μl
10×KBuffer2μl
0.1%BSA2μl
DNA3ul
灭菌水upto20μl37℃4h电泳1h
期待着高手们的指点,谢谢。
【之前构建的4个重组质粒里也有两个出现这样的问题(aagtcc变成agtcc),但是酶切能切出插入的片段】
目前问测序公司得到的答复是样品可能不是单克隆,测序结果是不准确的,建议重新挑取单克隆测序。
不知道还有没有别的方法?
●应用酶水解多肽不会破坏氨基酸,也不会发生消旋化。水解的产物为较小的肽段。向左转|向右转

