• 产品编号:NADPH 100-2/-3
    • 产品名称:Fluoro NADP™ Fluorescent NADP/NADPH Detection Kit - Detection of NADP/NADPH in tissue extracts /cell samples/荧光NADP/NADPH检测试剂盒
    • 产品规格:100Test/500Test
    • 价  格:7425.00/35625.00
    • 供 应 商:Cell Tech
    • 库  存:
  • Fluoro NADP™ NEW!
    Fluorescent NADP/NADPH Detection Kit - Detection of NADP/NADPH in tissue extracts /cell samples
     
    Key Benefits:
    • Detection of NADP/NADPH activity in cells or tissue extracts
    • Detection of NADP/NADPH levels in antioxidation and oxidative stress
    • Detection of NADP/NADPH levels in cell death, energy metabolism, and mitochondria function
    • Species Independent - NADP/NADPH detection in Bacterial, fungal and plant cells
    • Highly Sensitive – Detects up to 4nM NADP and NADPH
    • Highly Specific - No Cross reactivity with NAD/NADH
    • Easy to Use – 96 well Fluorescent Plate reader readout
       
    Introduction
    Nicotinamide adenine dinucleotide phosphate (NADP+) is used in anabolic reactions, such as lipid and nucleic acid synthesis, which require NADPH as a reducing agent. NADPH is the reduced form of NADP+, and NADP+ is the oxidized form of NADPH. In cells, NADPH plays the role of a carrier of reducing power and is primarily involved in maintaining optimal redox metabolism. A simplified assay for the measurement of NAD and NADP is critical to understanding the roles of these pyridine nucleotides in normal and abnormal cells.

    NADPH is produced in the oxidative phase of the pentose phosphate pathway in cells, a multifunctional pathway whose primary purpose is to generate reducing power in the form of NADPH. NADPH is a cofactor for enzymes that synthesize energy-rich molecules and provide the reducing equivalents for the oxidation-reduction involved in protecting the cell from the toxicity of reactive oxygen species (ROS) and NADPH oxidase-dependent ROS generation. Both NAD and NADP have been shown to influence hemoglobin affinity for oxygen in erythrocytes. In plant cells, NADPH is used as the reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis (1-2).

    Cell Technology’s Fluoro NADP/NADPH provides a highly reliable, sensitive fluorometric assay for the quantification of NADP, NADPH and their ratio in biological samples.
     
    Assay Principle
    The Fluoro NADP/NADPH detection kit utilizes a non-fluorescent detection reagent, which is oxidized in the presence NADPH to produce its fluorescent analog and NADP. NADP is further converted to NADPH via an enzyme-coupled reaction. The enzyme reaction specifically reacts with NADP/NADPH and not with NAD/NADH.

    Reaction:

    1. NADPH + non-fluorescent detection reagent + electron coupler → fluorescent analog + NADP

    2. NADP + enzyme coupled reaction →  NADPH (then proceeds to reaction 1).

       Excitation: 530-570nm and Emission at 590-600nm
     
    References
    1. Comprehensive Invited Review WEIHAI YING. NAD /NADH and NADP /NADPH in Cellular Functions and Cell Death: Regulation and Biological Consequences. ANTIOXIDANTS &REDOX SIGNALING Volume 10,Number 2,2008.

    2. Bedard K and Krause KH. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology.Physiol Rev 87:245 –313,2007.

    3. Lowry, Oliver H., Passonneau, Janet V. and Rock, Martha K. The Stability of Pyridine Nucleotides. The Journal of Biological Chemistry, 236, #10, 1961.
    Kit contents (for 100 assays)
    1. Enzyme Mix 170μL                          Part # 6022 .......................... 4-8C 

    2. NADP/NADPH Lysis Solution 12.5mL   Part# 3050 ............................4-8C

    3. NADP Extraction Buffer 22 mL           Part# 3046 (Green label) ..........4-8C

    4. NADPH Extraction Buffer 22 mL         Part# 3047 (Magenta label)........4-8C

    5. Reaction Buffer 11 mL.                    Part# 3049 ............................4-8C

    6. Standard curve diluent 15 ml.            Part # 3052............................4-8C

    7. NADPH Detection reagent 110 μL       Part # 4019...........................-20C 

    8. 3X Substrate Mix 6mL                     Part # 3048........................... -20C 

    9. NADPH Standard- 3 vials dried          Part# 7014.............................-20C
     
     

    Fluoro NADP Fluorescent NADP/NADPH Detection Kit - Detection of NADP/NADPH in tissue extracts /cell samples/荧光NADP/NADPH检测

    荧光NADP/NADPH检测试剂盒

    检测组织提取物,细胞样品中荧光NADP/NADPH

     

    主要优点:

    检测细胞或者组织提取物中的NADP/NADPH的活性

    检测氧化抑制和氧化作用中NADP/NADPH的水平

    检测细胞死亡,能量新陈代谢,线粒体功能中NADP/NADPH的水平

    种属特异性---检测细菌,真菌,植物细胞中的NADP/NADPH

    高灵敏性--- 可以检测4nM NAD/NADP

    高特异性--- NAD/NADH没有交叉反应

    使用方便--- 96孔荧光酶标仪

     

    介绍:

    烟酰胺腺嘌呤二核苷酸磷酸(NADP+)用于合成代谢反应,如脂质和核酸的合成,这需要NADPH作为还原剂。 NADPHNADP +的还原形式,NADP +NADPH的氧化形式。在细胞中,NADPH作为一个载体,起到降低功耗的作用,主要用来参与保持****的氧化还原代谢水平。用于NADNADP的简化测量法,对于了解在正常和不正常的细胞中的吡啶核苷酸的作用是至关重要的。

     

    NADPH是在细胞的磷酸戊糖途径的氧化阶段产生的,磷酸戊糖途径是一个多功能的途径,其主要目的是产生NADPH,减少能耗。 NADPH是酶的一种辅助因子,用来合成能量丰富的分子以用于供给氧化还原反应失去的能量。NADNADP都已经被证明能够影响红细胞中血红蛋白氧亲和力。在植物细胞中,NADPH用于光合作用(1-2)卡尔文循环的合成反应中,降低能耗。

     

    Cell Technology的荧光NADP/NADPH提供了高可靠性,灵敏的荧光检测,用于NADPNADPH和它们在生物样品中的比率的荧光定量检测分析。

     

    检测原理:

    荧光NADP / NADPH检测试剂盒采用了非荧光检测试剂,在NADPH存在时可氧化产生荧光类似物和NADP NADP通过酶偶联反应可进一步转化为NADPH。该酶反应可以与NADP/ NADPH特异性反应,不与NAD / NADH反应。

     

    反应:

    1. NADPH + 非荧光检测试剂+ 电子偶联剂 荧光类似物 + NADP

     

    2. NADP + 酶偶联反应   NADPH (然后接反应1).

     

    激发波长: 530-570nm    发射波长: 590-600nm