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Citrate Synthase Activity Colorimetric Assay Kit

based on 2 citations in multiple journalsCitrate Synthase Activity Colorimetric Assay Kit24.1 4
Catalog #: K318
$450.00

Product Details

Cat # +Size K318-100
Size 100 assays
Detection Method Absorbance (412 nm)
Applications Measurement of citrate synthase activity in various tissues/cells
Analysis of intact mitochondria
Features & Benefits • Simple, rapid & convenient
• The assay can detect Citrate Synthase Activity less than 1 mU in a variety of samples
Kit Components • CS Assay Buffer
• CS Substrate Mix (Lyophilized)
• CS Developer (Lyophilized)
• GSH Standard (reduced) (Lyophilized)
• CS Positive Control (Lyophilized)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Citrate Synthase (EC 2.3.3.1) is a key enzyme that is present in all living organisms. It catalyzes the conversion of acetyl-CoA and oxaloacetate into citrate and serves as a marker for intact mitochondria. A recent study showed that increased activity of mitochondrial Citrate Synthase is directly associated with metabolic and endocrine abnormalities such as obesity. In BioVision’s Citrate Synthase Activity Assay Kit, Citrate Synthase reacts with substrate mix to form an intermediate, which subsequently reacts with developer to generate the colored product. The rate of color development is proportional to the enzyme activity. The assay is simple, rapid and can detect Citrate Synthase activity less than 1 mU in a variety of samples.


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We have already homogenized our mice tissues (skeletal muscle) with TNS buffer (25mM Tris-HCl/50mM NaCl/250mM Sucrose/20mM NaF/1mM DTT/1uM PMSF), and we are hoping that we can use this tissue lysate for citrate synthase activity assays. Do you think we can use our lysates for this assay?
DTT and PMSF might not be ideal for these assays. DTT is a reducing agent which can interfere in the detection reaction. We recommend using the kit assay buffer which have been optimized to work in these assays. If you have no option but to use the current sample, limit the sample volume in each well to <20% of the total reaction and fill up the volume with the assay buffer to minimize the effect of DTT.
Can this kit be used for mouse samples?
K318 assays for the activity of the citrate synthase enzyme. This is not species-specific. This assay can work in any sample which has active citrate synthase in it including mouse samples.
Can it be used on frozen material (snap frozen, stored at -80)?
Yes, this kit can be used with snap-frozen tissue stored at -80C. We always prefer fresh samples over frozen but is tissue is frozen right after collection this should work.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
In this kit, after mitochondria has been isolated, does the mitochondria get lysed as part of the reaction mix to determine Citrate Synthase activity within?
Yes, the assay buffer we have has detergent in it and it will lyse the mitochondria. Citrate synthase, although translated in the cytoplasm, is transported to the mitochondria where it is localized in the inner mitochondrial membrane and therefore the membrane needs to be lysed.
Annunziata N. Crupi, Oxidative muscles have better mitochondrial homeostasis than glycolytic muscles throughout life and maintain mitochondrial function during aging. Aging (Albany NY)., Nov 2018;  30449736.
Feilong Wang, et al., (2018) Interferon Gamma Induces Reversible Metabolic Reprogramming of M1 Macrophages to Sustain Cell Viability and Pro-Inflammatory Activity, EBioMedicine, Feb. 2018, 29463472
Wang, B. et al., (2017) Exercise restores bioavailability of hydrogen sulfide and promotes autophagy influx in livers of mice fed with high-fat diet, Canadian Journal of Physiology and Pharmacology, Jan.2017, 95(6), 667-674
Liu et al., Sequential Actions of SIRT1-RELB-SIRT3 Coordinate Nuclear-Mitochondrial Communication during Immunometabolic Adaptation to Acute Inflammation and Sepsis. J. Biol. Chem., Jan 2015; 290: 396 - 408.
For more citations of this product click here