This kits takes our stand alone assay for phospholipase A2, PLA2 (A10072) and combines the necessary Ez and lipids toRead more
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Catalog Number
Quantity
E10217
1 Kit
Catalog number E10217
Price (EUR)
459,00
Each
Add to cart
Quantity:
1 Kit
Price (EUR)
459,00
Each
Add to cart
This kits takes our stand alone assay for phospholipase A2, PLA2 (A10072) and combines the necessary Ez and lipids to run 2 complete 96 well microplate assays that monitor activity in purified enzyme preparations and cell lysates. The EnzChek Phospholipase A2 Kit provides enough reagents for 2 microplates, using 200 μl volumes in 96 well format to perform continuous fluorometric monitoring of PLA2 . This product offers an alternative to our (B7701), (bis-BODIPY® FL C11-PC) reagent, by providing an PLA2 selective substrate and one that is ratiometric, thereby lowering variations produced by instrumentation and assay conditions. Phospholipase A2 or PLA2 represents a family of enzymes that hydrolyze the sn-2 ester linkage of phospholipids. The activities of these enzymes play important roles in cardiovascular, inflammatory and nervous system disorders, and in cancers. The EnzChek® Phospholipase A2 substrate provides sensitive and continuous rapid real-time monitoring of PLA2 enzyme activities. This unique substrate is selective for PLA2 and can be used either in a intensity or ratiometric based detection mode. In intensity based detection mode the PLA2 activity is monitored by the intensity increase of a single wavelength at approximately 515 nm. In ratiometric analysis, which is based on the distinct fluorescence resonance energy transfer (FRET) emission of this substrate prior to and after cleavage, PLA2 is detected by changes in the emission intensity ratio at 515/575 nm with excitation at ≈ 460 nm. Either detection mode provides a simple method with low background, high sensitivity and high specificity for PLA2.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence Intensity
Dye TypeBODIPY™ FL
Filter TypeFITC (Fluorescein) Filter
Quantity1 Kit
Shipping ConditionRoom Temperature
Substrate PropertiesLipid-Based Substrate, Chemical Substrate
Substrate TypePhospholipase Substrate
Target EnzymePhospholipase
ColorMixture, Green
EmissionRatiometric: 460 to 490/515, 575, Intensity-based: 460 to 490/515
For Use With (Application)Phospholipase Assay
For Use With (Equipment)Microplate Reader
Product LineEnzChek
Product TypeEnzChek Assay Kit
Unit SizeEach
Contents & Storage
1 kit. Store kit at -<20°, protected from light, desiccated
Frequently asked questions (FAQs)
What is the LoD for the EnzChek Phospholipase A2 Assay Kit (Cat. No. E10217)?
The EnzChek Phospholipase A2 Assay Kit (Cat. No. E10217) can detect PLA2 at 0.05 U/mL or lower.
The FRET-based substrate Red/Green BODIPY PC-A2 shows no change in fluorescence upon enzymatic cleavage. I'm not seeing an increasing green signal with higher concentrations of phospholipase A2 and no change in red fluorescence. Why is this?
There are numerous factors that can interfere with FRET-based detection. Here are a few items to consider:
a) Is there any component in the reaction solution that can quench green fluorescence?
b) Is there any component in the reaction solution that autofluoresces in the green or red emission range?
c) Too high of a substrate concentration may result in diffusional FRET: Lower the final working concentration of the substrate.
d) Ensure adequate dispersal of the substrate during the reaction. BODIPY dyes are very hydrophobic, and aggregates of the green product, BODIPY FL, can form an excimer and emit in the red range.
You provide Phospholipase A2 from honey bee venom in the EnzChek Phospholipase A2 Assay Kit. Can the kit be used with phospholipase A2 from other species?
Yes, phospholipase A2 from other species may be used as long as the phospholipase A2 can hydrolyze the sn-2 ester linkage of phospholipids and the enzymatic reaction can be done within a pH of >3 and <9.
Crystal structure of the predicted phospholipase LYPLAL1 reveals unexpected functional plasticity despite close relationship to acyl protein thioesterases.
Authors:Bürger M, Zimmermann TJ, Kondoh Y, Stege P, Watanabe N, Osada H, Waldmann H, Vetter IR,
Journal:J Lipid Res
PubMed ID:22052940
'Sequence homology indicates the existence of three human cytosolic acyl protein thioesterases, including APT1 that is known to depalmitoylate H- and N-Ras. One of them is the lysophospholipase-like 1 (LYPLAL1) protein that on the one hand is predicted to be closely related to APT1 but on the other hand might ... More
Alterations in cerebrospinal fluid glycerophospholipids and phospholipase A2 activity in Alzheimer's disease.
Authors:Fonteh AN, Chiang J, Cipolla M, Hale J, Diallo F, Chirino A, Arakaki X, Harrington MG,
Journal:
PubMed ID:23868911
'Our aim is to study selected cerebrospinal fluid (CSF) glycerophospholipids (GP) that are important in brain pathophysiology. We recruited cognitively healthy (CH), minimally cognitively impaired (MCI), and late onset Alzheimer''s disease (LOAD) study participants and collected their CSF. After fractionation into nanometer particles (NP) and supernatant fluids (SF), we studied ... More
Group V secretory phospholipase A2 plays a pathogenic role in myocardial ischaemia-reperfusion injury.
Authors:Yano T, Fujioka D, Saito Y, Kobayashi T, Nakamura T, Obata JE, Kawabata K, Watanabe K, Watanabe Y, Mishina H, Tamaru S, Kugiyama K,
Journal:Cardiovasc Res
PubMed ID:21169294
Group V secretory phospholipase A(2) (sPLA(2)-V) is highly expressed in the heart. This study examined (i) the role of sPLA(2)-V in myocardial ischaemia-reperfusion (I/R) injury and (ii) the cooperative action of sPLA(2)-V and cytosolic PLA(2) (cPLA(2)) in myocardial I/R injury, using sPLA(2)-V knockout (sPLA(2)V(-/-)) mice. Myocardial I/R injury was created ... More