Lipid Radical Probe -NBD-Pen-

Lipid Radical Probe -NBD-Pen-

Lipid Radical Detection

  • Enables detection of intracellular lipid radicals
  • Compatible with fluorescence microscopy imaging and flow cytometric detection and quantification
  • Product code
    L272　 Lipid Radical Probe -NBD-Pen-
Unit size Price Item Code
10 nmol×1 Find your distributors L272-10
10 nmol×5 Find your distributors L272-12

*10 nmol can be used for 5 assays in 35 mm dish (final concentration 1 µmol/l).

The Importance of Lipid Radicals in Ferroptosis

Ferroptosis is a form of cell death triggered by iron-dependent lipid peroxidation and has been increasingly implicated in various diseases, including cancer, neurodegenerative diseases, and age-related disorders. Lipid radicals are key intermediates involved in the propagation of lipid peroxidation chain reactions and are attracting attention as indicators of early changes and progression of lipid oxidation during ferroptosis.

NBD-Pen enables detection of lipid radicals generated within cells. Simply add the reagent to cultured cells to detect intracellular lipid radicals using fluorescence microscopy or flow cytometry.

 

 

This product was developed under the guidance of Professor Kenichi Yamada of Kyushu University.

Product Name Target Detection Properties
FerroOrange Intracellular Fe2+ Microscopy, Plate reader
Ex: 543 nm / Em: 580 nm
Mito-FerroGreen Mitochondrial Fe2+ Microscopy
Ex: 505 nm / Em: 535 nm
Lyso-FerroRed Lysosomal Fe2+ Microscopy, FCM, Plate reader
Ex: 551 nm / Em: 571 nm
Iron Assay Kit -Colorimetric- Fe2+ and Fe3+ Plate reader
Colorimetric, λ: 593 nm
Liperfluo Lipid Peroxide Microscopy, FCM
Ex: 488 nm / Em: 500-550 nm
Lipid Peroxidation Probe
-BDP 581/591 C11-		 Lipid Peroxidation
Process

Microscopy, FCM, Plate reader
Pre-reaction, Ex: 488 nm / Em: 510-550 nm
Post-reaction, Ex: 561 nm / Em: 600-630 nm
MDA Assay Kit Malondialdehyde Plate reader
Fluorescence, Ex: 540 nm / Em: 590 nm
Colorimetric, λ: 532 nm
Cystine Uptake Assay Kit Cystine uptake Plate reader
Ex: 485 nm / Em: 535 nm
GSSG/GSH Quantification Kit II GSSG and GSH Plate reader
Colorimetric, λ: 405 nm

Manual

Technical info

Please select the appropriate reagent based on your experimental method and detection instrument used for lipid peroxidation analysis.

  Liperfluo Lysosomal Lipid Radical Probe -Lyso-NBD-Pen- [This Product] Lipid Radical Probe -NBD-Pen- Lipid Peroxidation Probe -BDP 581/591 C11-
Target Lipid peroxides Lipid radicals Lipid radicals Lipid radicals + hydroxyl radicals

Localization

 Intracellular

Lysosomal

Intracellular

 Intracellular

Compatible Instruments

Fluorescence microscopy,
​Flow cytometry

Fluorescence microscopy,
​Flow cytometry

 Fluorescence microscopy,
Flow cytometry

Fluorescence microscopy,
Flow cytometry,
Plate reader

Detection Conditions
(Fluorescence Microscopy)

Fluorescence
Ex: 488 nm, Em: 500–550 nm

 Fluorescence
Ex: 488 nm, Em: 490–600 nm		 Fluorescence
Ex: 488 nm, Em: 490–600 nm

Fluorescence
Green: Ex: 488 nm, Em: 510–550 nm
Red: Ex: 561 nm, Em: 600–630 nm
Sensitivity ☆ ☆☆ ☆☆☆

Interaction with Target

Reacts only
(does not bind)

 

Reacts and binds

 Reacts only
(does not bind)
Features Specifically detects lipid peroxides.

Binds to lipid radicals, enabling detection of changes in lipid localization after the reaction.

 Ratiometric detection enables quantification using a plate reader.

High Selectivity for Lipid Radicals

The reactive oxygen species listed below were mixed with this reagent in 100 mmol/l phosphate buffer (pH 7.4), and the fluorescence intensity was measured after a 30-minute reaction.

Detection of Intracellular Lipid Radicals

Treatment of HT1080 cells with the ferroptosis inducer RSL3 resulted in an increase in intracellular lipid radicals. Furthermore, co-treatment with the lipid radical scavenger Liproxstatin-1 (Lip-1) resulted in a decrease in the signal, indicating that this reagent selectively detects lipid radicals.


<Detection wavelength>
Ex/Em = 488/490–600 nm

Quantification of Intracellular Lipid Radicals

HT1080 cells were treated with the ferroptosis inducer RSL3, and the increase in intracellular lipid radicals was quantified by flow cytometry.

Fluorescence Properties

Q & A

Q

How many samples can be measured with this product?

A

The number of samples that can be measured with 10 nmol of this product is as follows:

  • 96-well plate: 1 plate
  • ibidi 8-well plate: 50 wells
  • 35 mm dish: 5 dishes
  • 6-well plate: 5 wells

Q

Is there an experimental example that can be used as a positive control?

A

Please refer to the instruction manual for the lipid radical detection experiment using HeLa cells treated with cumene hydroperoxide.

Q

Can NBD-Pen be used for observation after cell fixation following staining?

A

Yes. We have confirmed that cells can be observed after fixation with 4% paraformaldehyde in PBS following staining.

Q

What precautions should be taken when observing samples with a fluorescence microscope?

A

NBD, the fluorescent scaffold of this probe, is susceptible to photobleaching when continuously exposed to excitation light. When acquiring imaging data, adjust the focus under bright-field observation before acquiring fluorescence images.

Q

How long can samples be observed after staining?

A

We have confirmed that samples can be observed for up to 24 hours after staining.

Handling and storage condition

Specification
Appearance： Orange to red solid
Solubility in Dimethyl sulfoxide： To pass test
Dye content： To pass test
Handling and storage condition
Store at room temperature.
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