- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Key Benefits
- Sensitive Fluorescent Assay.
- Enzyme Positive Control included in kit.
- Can monitor multiple time points to follow kinetics.
- One-step, No Wash assay.
- Adaptable for High Throughput format.
- Monitors enzymatic activity.
- Applications – Fluorescent Plate Reader.
Additional information
| Kit Size | 500 |
|---|
Catalase is an antioxidant enzyme that catalyses the decomposition of hydrogen peroxide (H202) to water and oxygen. Catalase is ubiquitously expressed in mammalian and non-mammalian aerobic cells containing the cytochrome system. The enzyme has been isolated from various sources, including bacteria and plant cells (1-3). Catalase activity varies greatly from tissue to tissue. Highest activity is seen in liver and kidney, while lowest activity is seen in connective tissue (3). In eukarotic cells, catalase in concentrated in organelles called peroxisomes (4).
The production of hydrogen peroxide in eukaryotic cells is an end product result of various oxidases and superoxide dismutase reactions. Accumulation of H202 can result in cellular damage through oxidation of proteins, DNA and lipids thus resulting in cell death and mutagenisis (8-11). H202 role in oxidative stress related diseases have been widely studied (8,12).
The Fluoro Catalase detection kit is sensitive assay that utilizes a non – fluorescent detection reagent to detect H202 substrate left over from the catalase reaction (5-6).
| Document Title |
| Fluoro Catalase Protocol |
| Fluoro Catalase Datasheet |
| msds.FluoroCatalase |
| Title | File | Link | Author(s) | Journal | Year; Edition:Pages |
| Expression of HIF prolyl hydroxylase isozymes in growth plate chondrocytes: Relationship between maturation and apoptotic sensitivity | http://onlinelibrary.wiley.com/doi/10.1002/jcp.20873/full | S.P. Terkhorn, J. Bohensky, I.M. Shapiro, E. Koyama, V. Srinivas | Journal of Cellular Physiology | Volume 210, Issue 1, pages 257–265, January 2007- DOI: 10.1002/jcp.20873 | |
| Lethal and sublethal effects of the pyrethroid, bifenthrin, on grass shrimp (Palaemonetes pugio) and sheepshead minnow (Cyprinodon variegatus) | http://www.informaworld.com/smpp/content~db=all~content=a795388224 | Heather E. Harper; Paul L. Pennington; Jennifer Hoguet; Michael H. Fulton | Journal of Environmental Science and Health | Part B: Pesticides, Food Contaminants, and Agricultural Wastes Volume 43, Issue 6, 2008, Pages 476 – 483 DOI: 10.1080/03601230802174599 | |
| 3β-Hydroxysteroid-{Delta}24 Reductase Is a Hydrogen Peroxide Scavenger, Protecting Cells from Oxidative Stress-Induced Apoptosis | http://endo.endojournals.org/cgi/content/abstract/149/7/3267 | Xiuli Lu, Fukushi Kambe, Xia Cao, Yasuko Kozaki, Takahide Kaji, Takehisa Ishii and Hisao Seo | Endocrinology | Vol. 149, No. 7 3267-3273, 2008 | |
| Regulation of autophagy in human and murine cartilage: Hypoxia-inducible factor 2 suppresses chondrocyte autophagy | http://onlinelibrary.wiley.com/doi/10.1002/art.24444/full?globalMessage=0 | Jolene Bohensky1, Shawn P. Terkhorn1, Theresa A. Freeman1, Christopher S. Adams1, Joseph A. Garcia2, Irving M. Shapiro1, Vickram Srinivas | Arthritis and Rheumatism | Vol 60, Issue 5, pp 1406-1415, May 2009. DOI: 10.1002/art.24444 |
| Reference |
| Deisseroth, A., and Dounce, A.L. Catalase: Physical and chemical properties, mechanism of catalysis, and physiological role Physiol. Rev., 50, 319-375 (1970). |
| Sebastian Mueller, Hans-Dieter Riedel and Wolfgang Stremmel. Determination of Catalase Activity at Physiological Hydrogen Peroxide Concentrations. Analytical Biochemistry Volume 245, Issue 1 ,1 February 1997, Pages 55-60. |
| Deisseroth, A., and Dounce, A.L. Catalase: Physical and chemical properties, mechanism of catalysis, and physiological role. Physiol. Rev., 50, 319-375 (1970). |
| Marcel Zámocký and Franz Koller . Understanding the structure and function of catalases: clues from molecular evolution and in vitro mutagenesis. Progress in Biophys. Mol. Biol., 72, 19-66 (1999). |
| Mingjie Zhou, Zhenjun Diwu, Nataliya Panchuk-Voloshina and Richard P. Haugland. A Stable Nonfluorescent Derivative of Resorufin for the Fluorometric Determination of Trace Hydrogen Peroxide: Applications in Detecting the Activity of Phagocyte NADPH Oxidase and Other Oxidases. Anal Biochem 253, 162 (1997). |
| J. G. Mohanty, Jonathan S. Jaffe, Edward S. Schulman and Donald G. Raible. A highly sensitive fluorescent micro-assay of H202 release from activated human leukocytes using a dihydroxyphenoxazine derivative. J. Immunol Methods 202, 133 (1997). |
| Tatyana V.Votyakova ,Ian J.Reynolds. Detection of hydrogen peroxide with Amplex Red:interference by NADH and reduced glutathione auto-oxidation. Archives of Biochemistry and Biophysics, 431:138-144 (2004). |
| Jingxiang Bai, Ana M. Rodriguez, J. Andres Melendez, and Arthur I. Cederbaum. Overexpression of Catalase in Cytosolic or Mitochondrial Compartment Protects HepG2 Cells against Oxidative Injury. J. Biol. Chem., Sep 1999; 274: 26217 - 26224 |
| Tada-Oikawa, S. et al., FEBS Lett., 442, 65-69 (1999). |
| Hampton, M.B., and Orrenius, S., FEBS Lett., 414, 552-556 (1997). |
| Kowaltowski, A.J. et al., FEBS Lett., 473, 177-182 (2000). |
| Tome, M.E. et al., Cancer Res., 61, 2766-2733 (2001). |
| Part# | Reagent | Temperature |
| 3021 | 5X Reaction Buffer pH 7.4, 25mL | 2-8C |
| 4010 | Detection Reagent, 1 Vial | 2-8C - Undiluted; Diluted: -20C(Aliquot in Single Use Vials) |
| 6009 | Horseradish Peroxidase, 1 Vial | 2-8C - Undiluted; Diluted: -20C |
| 3022 | Hydrogen Peroxide, 1 Vial | 2-8C |
| 6008 | Catalase Enzyme, 1 Vial | 2-8C (Crystalline Suspension) |
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反应体系如下:
plasmid10ul
10xKbuffer5ul
KpnI1ul
BamHI1ul注意千万不可多加总酶量必须<4%
ddH20upto50ul
总体积改变加酶量按比例改变.
因为溶蛋白酶是蛋白质本质的酶
如果能分解,那小肠液中的消化酶如何大量共存,如果不能
那它如何识别其他蛋白质物质是不是消化酶?
炎性体(inflammasome)是细胞内的一类多蛋白复合物,在炎性反应中发挥着至关重要的作用。炎性体包括半胱天冬酶-1(caspase1)、PYCARD和NALP,有时也包括半胱天冬酶-5(caspase5,也被称作半胱天冬酶-11或ICH-3)。它是在骨髓细胞(myeloidcell)中产生的,也是先天性免疫系统的一个组分。炎性体的确切组成依赖于启动炎性体组装的激活物,如双链RNA和石棉会引发不同的炎性体组成。炎性体促进炎性细胞因子IL-1β和IL-18成熟。
在一项新的研究中,来自比利时法兰德斯生物技术中心/根特大学(VIB/UGent)的LieselotteVandeWalle博士、DanielJiménezFernández以及教授MoLamkanfi研究团队对半胱天冬酶-12(caspase12)的功能产生新的认识。基于此,他们打破了这个领域对半胱天冬酶-12的固执观念:半胱天冬酶-12是炎性体的负调节物。这些新的认识为研究人员挣脱现有的研究路线和鉴定它的真正生理学功能铺平道路。相关研究结果发表在2016年6月2日那期Nature期刊上,论文标题为“Doescaspase-12suppressinflammasomeactivation?”。
研究人员也指出这将需要进行大量的“重新研究(re-researching)”。之前所谓的半胱天冬酶-12在细胞死亡、应激反应、疟疾和败血症等中的作用---引用了9000多次---必需复核,这是因为这些作用经常是基于不正确的小鼠模型得出的。
VIB/UGent教授MoLamkanfi说,“我们发现在很多情形下,对半胱天冬酶-12的研究是基于对半胱天冬酶-11(caspase11)和半胱天冬酶-12都进行基因敲除的小鼠模型开展的。因此从研究结果中推断半胱天冬酶-12的作用是不可能的。我们如今引入新的选择性半胱天冬酶-12基因敲除小鼠,这应当能够让我们追踪半胱天冬酶-12的确切功能。”
利用这些新的小鼠,Lamkanfi团队证实在体外模拟的BMDM(bonemarrow-derivedmacrophage,骨髓衍生性巨噬细胞)和体内接种的小鼠中,半胱天冬酶-12缺乏都不能增加半胱天冬酶-1激活。剔除半胱天冬酶-12也不会增强炎性体途径释放出成熟的IL-1β和IL-18。他们的发现表明不论半胱天冬酶-11的表达状态如何,半胱天冬酶-12都不会作为半胱天冬酶-1激活的生理学上负显性调节物,因而也不会作为炎性体的生理学上负显性调节物。

