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

Catalog No.	Quantity
A10072	100 μg

Catalog No. A10072 Supplier Invitrogen™ Supplier No. A10072

Description

Measure results in microplate assays or using flow cytometry

Molecular Probes phospholipase substrates are dye labeled phospholipids used to monitor phospholipase A (PLA) activity in purified enzyme preparations, cell lysates, and living cells. Two structural variations of these substrates are available, which confer specificity for PLA1 and PLA2. Fluorescently labeled versions of the cleavage product generated from these substrates are also available for use as standards for evaluating percentage substrate conversion in enzymatic reactions.

• Label (Ex/Em): Red/Green BODIPY™ PC-A2 (approx. 488/530,590nm) (can be used in dual emission fluorescence ratio detection)
• Specificity: PLA₂
• Cleavage product: BODIPY FL C5
• Lyophilized product can be dissolved in DMSO for use
• Fluorescence is measured either in microplate assays or using flow cytometry

Enzyme and Protein Activity Assays, Lipase and Phopholipase Activity, Proteins, Expression, Isolation and Analysis

Order Info

Shipping Condition: Room temperature

Specifications

• Cell Permeability: Cell-Permeant
• Content And Storage: Store in freezer (-5 to -30°C).
• Detection Method: Fluorescence
• Form: Solid
• Product Line: BODIPY
• Quantity: 100 μg
• Substrate: Phospholipase A2 substrate (ratiometric)
• Substrate Properties: Lipid-Based Substrate
• Target Enzyme: Phospholipase
• For Use With (Equipment): Flow Cytometer, Microplate Reader
• Shipping Condition: Room Temperature

Frequently Asked Questions (FAQs)

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.

For Research Use Only. Not for use in diagnostic procedures.