Skip to Content
Merck
All Photos(1)

Key Documents

View All Documentation

MAK122

Sigma-Aldrich

Phospholipid Assay Kit

sufficient for 100 colorimetric or fluorometric tests

Synonym(s):

Phospholipid Quantification Kit

Sign Into View Organizational & Contract Pricing
All Photos(1)

About This Item

UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

usage

sufficient for 100 colorimetric or fluorometric tests

application(s)

cosmetics
food and beverages
pharmaceutical

detection method

colorimetric
fluorometric

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

Phospholipids are a class of lipids which constitute a major component of cell membranes and play important roles in signal transduction. Most phospholipids contain one diglyceride, a phosphate group, and one choline.

In this assay, phospholipids (such as lecithin, lysolecithin and sphingomyelin) are enzymatically hydrolyzed to choline which is determined using choline oxidase and a H2O2-specific dye. This results in a colorimetric (570 nm)/ fluorometric (λex = 530/λem = 585 nm) product directly proportional to the phospholipid concentration in the sample. The range of linear detection is 3 - 200 μM for colorimetric assays and 0.6 - 20 μM for fluorometric assays.

Application

Phospholipid assay kit has been used in biochemical analysis[1] and phospholipid quantitation.[2][3][4][5]

Features and Benefits

Compatible with high-throughput handling systems.

Suitability

Suitable for the detection of phospholipids in biological samples

Principle

In this assay, phospholipids (such as lecithin, lysolecithin and sphingomyelin) are enzymatically hydrolyzed to choline which is determined using choline oxidase and a H2O2-specific dye. This results in a colorimetric (570 nm)/ fluorometric (λex = 530/λem = 585 nm) product directly proportional to the phospholipid concentration in the sample. The range of linear detection is 3 - 200 μM for colorimetric assays and 0.6 - 20 μM for fluorometric assays.

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334,H412

Precautionary Statements

P261 - P273 - P284 - P501

Hazard Classifications

Aquatic Chronic 3 - Resp. Sens. 1

Storage Class Code

10 - Combustible liquids

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
122CE11A20
122CE11A20
122CE08A29
122CD11A30
122CE08A29

Don't see the Right Version?

If you require a particular version, you can look up a specific certificate by the Lot or Batch number.

Search for a COA

Already Own This Product?

Find documentation for the products that you have recently purchased in the Document Library.

Visit the Document Library

Epithelial disruption of Gab1 perturbs surfactant homeostasis and predisposes mice to lung injuries.
Wang K, et al.
American Journal of Physiology. Lung Cellular and Molecular Physiology, 311(6), L1149-L1159 (2016)
Renoprotective effect of curcumin against the combined oxidative stress of diabetes and nicotine in rats.
Ibrahim ZS, et al.
Molecular Medicine Reports, 13(4), 3017-3026 (2016)
Prevention of gallbladder hypomotility via FATP2 inhibition protects from lithogenic diet-induced cholelithiasis.
Tharp KM, et al.
American Journal of Physiology: Gastrointestinal and Liver Physiology, 310(10), G855-G864 (2016)
Kevin M Tharp et al.
American journal of physiology. Gastrointestinal and liver physiology, 310(10), G855-G864 (2016-04-02)
Gallstone disease is a widespread disorder costing billions for annual treatment in the United States. The primary mechanisms underlying gallstone formation are biliary cholesterol supersaturation and gallbladder hypomotility. The relative contribution of these two processes has been difficult to dissect
Inhibiting mevalonate pathway enzymes increases stromal cell resilience to a cholesterol-dependent cytolysin.
Griffin S, et al.
Scientific Reports, 7(1), 17050-17050 (2017)
View All Related Papers

Questions

1–3 of 3 Questions  

  1. Is the 10 ml of Assay Buffer in MAK122 sufficient for 100 assays? Also, what amount of sample/buffer should be used?

    1 answer

    1. 1. The general recommendation for homogenization is a 1:10 solid to buffer ratio, for example, 20mg solid with 200 uL of Buffer. However, this ratio may not always be optimal. If there are concerns about having enough assay buffer for both homogenization and assays, the homogenization can be run with a different homogenization buffer, provided that it contains at least 0.5% Triton X-100. Please note that the amount of Assay Buffer needed per sample should be enough to yield 40 microliters of supernatant, unless you choose to use your own homogenization buffer with 0.5% Triton X-100.

      2. The kit is usually tested on tissue samples. However, it has not been tested on tissues themselves.

      3. For homogenizing samples, they can be run through 10-20 passes in a Dounce homogenizer on ice or by sonication, preferably performed in an ice-water bath. The degree of tissue lysis can be checked under a microscope. After homogenization, it is recommended to centrifuge the homogenate at 14,000 g for 10 minutes.

      Helpful?

  2. Why is cold assay buffer listed as a potential cause for the assay not working correctly? Can a 37 or 25 degree water bath be used to thaw the assay buffer? Can the assay buffer be stored at room temperature instead of at -20 degrees?

    1 answer

    1. The troubleshooting bulletin for MAK122 suggests that if the assay does not work, it could be due to the assay buffer not being at room temperature. This is because the enzyme is more active at room temperature than in colder conditions. Although stability data for the assay buffer at room temperature is not available, it is recommended to store it at -20 degrees or evaluate the suitability of room temperature storage. While the composition of the assay buffer is proprietary, it is expected to be stable at room temperature. Additionally, a 37 or 25 degree C water bath can be used to thaw the assay buffer, with a caution against excessive heating and a recommendation to remove it from the water bath once it reaches room temperature.

      Helpful?

  3. To prepare the 0.5% Triton X-100 solution to dilute my samples, should I dilute Triton x-100 in water, in the assay buffer, or something else?

    1 answer

    1. The 0.5% Triton X-100 solution to be used as the sample diluent may be prepared using water.

      Helpful?

Reviews

No rating value

Active Filters

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service