Citrate Synthase Activity Assay试剂盒(ab119692)

概述

  • 产品名称Citrate Synthase Activity Assay试剂盒
  • 检测方法Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    1 3 5.45%
    批次间
    样品 n Mean SD CV%
    1 3 8.3%
  • 样品类型
    Cell culture extracts, Tissue Extracts
  • 检测类型Enzyme activity
  • 灵敏度
    1.56 µg/ml
  • 范围
    1.56 µg/ml - 100 µg/ml
  • 实验步骤Multiple steps standard assay
  • 种属反应性
    与反应: Human
    不与反应: Mouse, Rat
  • 产品概述

    Citrate Synthase Activity Assay Kit (ab119692) is used to determine mitochondrial citrate synthase (CS) activity in a sample at an immunocapture based manner. The enzyme is captured within the wells of the microplate, and activity is determined by recording color development of TNB, which is generated from DTNB present in the reaction of citrate synthesis. The overall reation product, TNB, absorbs at OD = 412 nm.

    This kit specifically immunocaptures only native citrate synthase in well from the applied test sample. The immunocapture based activity assay allows researchers to measure citrate synthase activity with simple sample preparation and no need for mitochondria isolation.

    This kit contains all of the required reagents for a fast and simple measurement of citrate synthase activity in a whole cell extract or in tissue homogenate.

  • 说明

    Citrate synthase (CS, O75390) is the initial enzyme of the tricarboxylic acid (TCA) cycle. This enzyme is a 51.7 kDa enzyme that catalyzes the reaction of 2 carbon acetyl CoA with 4 carbon oxaloacetate to form the 6 carbon citrate (EC 2.3.3.1). This enzyme is an exclusive marker of the mitochondrial matrix. Citrate synthase is found in nearly all cells capable of oxidative metabolism.

    Storage: All components are shipped cold. 10X Reaction Reagent mixture is shipped lyophilized. Before use, rehydrate the reaction mixture by adding 1mL base buffer. Vortex the tube thoroughly to dissolve the lyophilized powder to prepare 10X Reaction solution. After hydration unused amounts of the 10X Reaction solution should be stored at -80°C for 6 months. Store all other components at 4°C. This kit is stable for 6 months from receipt.

  • 经测试应用适用于: Functional Studiesmore details
  • 平台Microplate

性能

应用

Our Abpromise guarantee covers the use of ab119692 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.

Citrate Synthase Activity Assay Kit 图像

  • Figure 1. Example activity measurements from titrated HepG2 cells.
  • Figure 2. Example activity measurements from titrated of Human heart homogenate.

实验方案

Citrate Synthase Activity Assay Kit (ab119692)参考文献

This product has been referenced in:
  • Marrache S  et al. Detouring of cisplatin to access mitochondrial genome for overcoming resistance. Proc Natl Acad Sci U S A 111:10444-9 (2014). Read more (PubMed: 25002500) »
  • Gu F  et al. Alterations in mitochondrial DNA copy number and the activities of electron transport chain complexes and pyruvate dehydrogenase in the frontal cortex from subjects with autism. Transl Psychiatry 3:e299 (2013). Read more (PubMed: 24002085) »

See all 3 Publications for this product

Product Wall

Abreviews
This kit was purchased to see if it was compatible with mitochondria isolated from bovine PBMCs. The initial data looks good but more tests are needed to see if bovine samples can be stored at -80C for 6 months like stated in the protocol booklet. The image I attached shows the following data: A1 B1 (Blank), C1 D1 (Fresh Sample A), E1 F1 (Fresh Sample B), G1 H1 (1 month old sample C). There was some variation in the methods used to isolate the mitochondria in the fresh and 1 month old sample as we are still working to improve this part of the experiment. It is unknown if the variation in mitochondria isolation methods or the age of the samples used is responsible for the differences observed graphically.
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提交于 Sep 27 2016

1. This kit passed QC test so all the components should be prepared properly.
2. Try using a higher protein concentration (e.g. 500ug/ml in lieu of 100ug/ml), since different cell lines have different expression level of CS.
3. Increase the i...

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Same process for converting all to molarity per amount of protein loaded into well.
Note these are all per cm and should be converted into path length for microplate which should be about 0.6cm.

Extinc...

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I have received word back from the lab regarding your request. Unfortunately the details of the reagents and reactions for these products are proprietary and we cannot share them. They have provided what detail...

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The composition of the extraction buffer (P/N 8201093) is Hepes, NaCl, and a detergent, pH 7.4. Thus, it does not contain any of the other components you are concerned about (Tris-HCl, KPO4, or EDTA).

...

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Thank you for contacting us. You may be interested in the following kits:

Citrate Synthase: ab119692
Aconitase: ab109712 or ab83459
CitrateLyase: wedo not currently have a kit available, but weyou may be interested in ourprimary ant...

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Yes you can this kit uses lauryl maltoside to extract active enzymes.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Us...

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I have heard back from the lab with the following information:

All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/micro...

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Unfortunately, this kit does not work in mouse. The capture antibody works in human but not in rat or mouse. Other species have not been tested yet.


This is the only Citrate Synthase Activity ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"