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Phosphatidylcholine Colorimetric/Fluorometric Assay Kit

based on 3 citations in multiple journalsPhosphatidylcholine Colorimetric/Fluorometric Assay Kit34.1 4
Catalog #: K576

In stock

$375.00

Product Details

Cat # +Size K576-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications BioVision’s Phosphatidylcholine kit measures PC in the range of 0.1 to 10 nmol per sample.
Features & Benefits • Simple procedure; takes less than ~40 minutes• Fast and convenient
Kit Components • PC Assay Buffer
• OxiRed Probe
• PC Hydrolysis Enzyme
• PC Development Mix
• PC Standard (10 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Phosphatidylcholine (PC) is a phospholipid which incorporates choline as the headgroup of the lipid. PC is a major constituent of biological membranes and is involved in cell signaling through release of choline by phospholipase D leaving the second messenger phosphatidic acid. BioVision's Phosphatidylcholine Assay Kit is a simple convenient means of measuring Phosphatidylcholine in a variety of biological samples. The kit utilizes an enzyme-coupled assay in which PC is hydrolyzed, releasing choline which is subsequently oxidized resulting in development of the OxiRed probe to generate fluorescence (Ex/Em 535 nm 587 nm) and absorbance (570 nm). BioVision’s Phosphatidylcholine kit measures PC in the range of 0.1 to 10 nmol per sample. PC is present in serum at ~ 0.2-2.5 mM (~50-200 mg/dL).


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Can you suggest a protocol for testing cell culture supernatant/media?
Here’s a general protocol for testing cell media with Kit K576-100: Grow cells in the medium of choice (cell number to be empirically decided). Preferably use medium without choline chloride and phenol red as these can add to the background. If its detrimental to cell growth, then you can try switching the media before assay or do a background correction with media only. Add 1-10ul of cell culture medium directly to the well and dilute to 50ul with assay buffer. You can add upto 20ul if needed. Proceed with the rest of the protocol.
Can phospholipase D also cleave the lysophosphatidylcholine in cell lysate?
Lysophosphatidylcholine is not a substrate for Phospholipase D. Although there is some evidence in the literature that it can stimulate PhospholipaseD .
What is the source of the Phospholipase D that comes in the PC Hydrolysis Enzyme?
The enzyme is from Streptomyces chromofuscus.
Is there an interference from sphingomyelin in the assay?
Phosphatidylcholine and sphingomyelin both have phosphocholine headgroups. Hence this could be the reason why there is a strong signal from pure sphingomyelin which could potentially have some ~2% contamination from choline. The phospho-choline head groups might react with the phospholipase and lead to release of choline and hence interfere in the assay.
What is the protocol for testing cell culture supernatant? How did you concentrate this?
PC is amphipathic and can form vesicles. Even if there is PC in the cell culture supernatant, the amount might not be enough to be detected from a maximum of 100,000 cells in culture. Another consideration is the choline background which should be subtracted from the final PC numbers with the hydrolysis enzyme.
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.
Yuyama et al., Decreased Amyloid-β Pathologies by Intracerebral Loading of Glycosphingolipid-enriched Exosomes in Alzheimer Model Mice. J. Biol. Chem., Aug 2014; 289: 24488 - 24498.
Mitsutake, S. et. al. Dynamic Modification of Sphingomyelin in Lipid Microdomains Controls Development of Obesity, Fatty Liver, and Type 2 Diabetes. J. Biol. Chem., Aug 2011; 286: 28544 - 28555.
Li, Y. et. al. Contribution of sams-1 and pmt-1 to lipid homoeostasis in adult Caenorhabditis elegans. J. Biochem., May 2011; 149: 529 - 538.
For more citations of this product click here
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