- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Key Benefits
- Non Radioactive.
- Can monitor multiple time points to follow kinetics.
- Monitors Enzymatic Activity.
- One-step, no wash assay.
- Adaptable for High Throughput format.
- Enzyme Positive Control included in Kit.
- Applications – Fluorescent Plate Reader.
Additional information
| Kit Size | 500 |
|---|
Semicarbazide-sensitive amine oxidase (SSAO) is a common name for a widely distributed enzyme in nature. In man this enzyme is present in the vascular system and circulates in plasma. SSAO differ from the monoamine oxidases A and B in substrate and inhibitor patterns. These enzymes have been widely studied and their tissue distribution, molecular properties, substrate specificities and inhibitor sensitivities are extensively reviewed (2,3). SSAO exists in two forms: tissue bound and soluble (plasma SSAO). Tissue bound SSAO acitvity is associated with blood vessels, mainly in smooth muscle layers, however it is also associated with spleen, placenta, bone marrow, kidney, sclera, retina, endothelial cells, adipocytes, chondrocytes and fibroblasts. (4,5). Evidence suggests that Plasma SSAO originates from the cleavage of membrane-bound form. The possible sources of plasma SSAO are still unclear, but it has been suggested that it may be derived from liver, retina, placenta and bone tissue (6,7,8). SSAO’s functional role has been suggested to be involved in: apoptosis, atherogenesis, cell adhesion, leucocyte trafficking, glucose transport and local production of hydrogen peroxide. Reports of elevated levels of SSAO have been reported in congestive heart failure, diabetes mellitus, alzheimer’s disease and various other inflammatory diseases (1). Furthermore byproducts of SSAO deamination, such as formaldehyde and methylglyoxal, have been proposed to be involved in pathogenesis of cancer, aging and atherosclerosis. The Fluoro SSAO detection kit utilizes a non- fluorescent detection reagent, to detect H202 released from the conversion of Benzylamine to Benzaldehyde via SSAO. Furthermore H202 oxidizes this detection reagent in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase.
| Document Title |
| SSAOProtocol |
| Fluoro SSAO Datasheet |
| msds.fluoroSSAO |
| Reference |
| Yayin Tahiri Kasim et.al., Turk J Biochem; 29 (3);247-254 (2004); Semicarbazide-Sensitive Amine Oxidase: Biochemical and Physiological Properties |
| B.A.Callingham,A.Holt,J.Elliott.J.Neural Transm.(Suppl.)32 ,279 –290 (1990). |
| B.A.Callingham and M.A.Barrand.J.Neural Transm.(Suppl.)23 ,37 –54 (1987). |
| G.A.Lyles.J.Neural Transm.(Suppl.)41 ,387 –396 (1994). |
| K.Magyar, Z.Mészáros and P.Mátyus . Semicarbazide-sensitive amine oxidase.Its physiological significance Pure Appl.Chem.,Vol.73,No.9,pp.1393 –1400,2001. |
| Boomsma F.,van Dijk J.,Bhaggoe U.M.,Bouhuizen A.M.B.,van den Meiracker A.H.(2000)Variation in semicarbazide-sensitive amine oxidase activity in plasma and tissues of mammals.Comp.Biochem. |
| Ekblom J.,Gronval J.L.,Garpenstrand H.,Nillson S.,Oreland L.(2000)Is semicarbazide-sensitive amine oxidase in blood plasma partly derived from the skeleton?.Neurobiology,8,129-135. |
| McGuirl M.A., Dooley D.M.(1999)Copper-containing oxidases. Curr.Opin.Chem.Biol.,3,138-144. |
| Methods Enzymology 142, 617 (1997). |
| 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). |
| Biochem Pharmacol 18, 1447 (1969). |
| Part# | Reagent | Temperature |
| Part # 3019 | 5X Reaction Buffer pH 7.4, 1 Bottle | 2-8C |
| Part # 6005 | Horseradish Peroxidase, 1 Vial | 2-8C |
| Part # 4008 | Detection Reagent, 1 Vial | -20C (Aliquot in Single Use Vials) |
| Part # 7001 | SSAO Substrate Benzylamine, 1 Vial | -20C (Aliquot in Single Use Vials) |
| Part # 6006 | SSAO Enzyme, 1 Vial | -20C (Aliquot in Single Use Vials) |
| Part # 7003 | Pargyline: Monoamine Oxidase B Inhibitor(9-11), 1 Vial | -20C (Aliquot in Single Use Vials) |
| Part # 7004 | Semicarbazide-Sensitive Amine Oxidase Inhibitor, 1 Vial | -20C (Aliquot in Single Use Vials) |
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DNA连接酶主要是连接DNA片段之间的磷酸二酯键最初从原核生物（大肠杆菌）分离得到的.现在生物基因工程主要是从T4噬菌体中分离得到的,
菌体构建时，目的基因连接在T载体上，测序也正确，但是将目的基因还有载体分别双酶切后总是连接不上，将连接后产物跑核酸胶，什么也没有，不知道是什么原因？我的目的基因浓度为9ng/ul,载体回收后的浓度为6ng/ul,是因为浓度太低的原因吗？还有就是连接有目的基因的T载体双酶切后出现了三条带，这个又是什么原因呢？希望得到解答
连接酶通常是包括“连接酶”这个字，就如DNA连接酶是将脱氧核糖核酸（DNA）片段连接。其他普遍的名称包括“合成酶”，因为这些酶是用作合成新的分子，或当它们是将二氧化碳加入一个分子时则称为“羧化酶”。
大家有用过Invitrogen的T4连接酶吗？说明书上是23－26度连接，一般的连接酶不都是16度吗？应该用多少度呢？另外，说明书上还说连接后为了达到更好的转化效率，应将连接反应液至少稀释5倍再转化，是这样吗？谢谢大家帮忙啊
只不过限制性核苷酸酶是将磷酸二酯键切断；而DNA连接酶则是形成磷酸二酯键。
求有经验的大神指教，我的载体和目的基因连不上，转化不到大肠中，比例为1:3。另外，为啥胶回收后的载体浓度那么低，大约6ng/ul了，影响连接么？
