Red/Green BODIPY™ PC-A2 (1-O-(6-BODIPY™ 558/568-Aminohexyl)-2-BODIPY™ FL C5-Sn-Glycero-3-Phosphocholine) (Ratiometric Phospholipase A2 Substrate) - FAQs

2 个常见问题解答

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.

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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.

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