Lipid Peroxidation Probe -BDP 581/591 C11-

Lipid Peroxidation Probe -BDP 581/591 C11-

Lipid Peroxidation Assay

  • High sensitivity to Lipid Peroxidation
  • Applicable to Fluorescent Microscope, Plate Reade, Flow Cytometer
  • Product code
    L267  Lipid Peroxidation Probe -BDP 581/591 C11-
Unit size Price Item Code
200 tests $280 L267-10
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200 tests
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Description

 Reactive oxygen species (ROS) are highly reactive chemicals formed from diatomic oxygen (O2). In recent years, it has been reported that ROS reacts with superoxide causing lipid peroxidation, which is associated with cell death and various diseases such as inflammation. As described above, ROS undergoes various mechanisms and therefore, it is important to evaluate the lipid peroxidation level with appropriate reagents.
 Lipid Peroxidation Probe -BDP 581/591 C11- is a fluorescent dye used to detect the formation of lipid peroxides.  Dojindo Laboratories' (Code: L248) Liperfluo can detect lipid peroxides. Unlike Liperfluo, This fluorescent dye does not respond to lipid peroxides but reacts with lipid radicals formed during lipid peroxidation by reactive oxygen species with high sensitivity.  The high sensitivity of this probe enables detection with a plate reader, which is difficult with Liperfluo, making it an excellent reagent for fluorescence quantification and screening applications.

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Manual

Technical info

 Lipid Peroxidation Probe -BDP 581/591 C11- is a fluorescent probe for detecting lipid peroxidation. This fluorescent probe does not react with lipid peroxides but reacts with lipid radicals generated when lipids are peroxidized, resulting in the detection of lipid peroxidation. The unreacted probe emits red fluorescence, but after reacting with radicals around lipids, it changes its fluorescence from red to green. Thus, lipid peroxidation can be detected with high sensitivity because it is detected by the ratio of red to green fluorescence intensity.


Spectrum of Lipid Peroxidation Probe -BDP 581/591 C11

Application Data: Lipid Peroxidation Assay of t-BHP treated HepG2 cells

HepG2 cells stained with this probe were stimulated with HBSS solution containing 200 µmol/l t-BHP for 2 hours, and the fluorescence intensity was compared with control cells. As a result, a decrease in red fluorescence and an increase in green fluorescence were observed with high sensitivity in t-BHP-treated cells compared to untreated cells. The cells were detected using a plate reader, and the values obtained were calculated as the intensity ratio of green/red fluorescence, which allowed quantified lipid peroxidation. Furthermore, an increase in the histogram of green fluorescence was observed when the cells were detected using a flow cytometer. It was found that this dye can be analyzed using three different instruments.


<Experimental Conditions>
Instrument: Fluorescent Microscope
Green: GFP filter (Ex = 450-490 nm, Em = 500-550 nm)
Red: TexasRed filter (Ex = 540-580 nm, Em = 600-660 nm)
Scale bar: 50 µm


<Experimatal Conditions>
Instrument: Fluorescent Plate Reader
Green: Ex = 490 nm, Em = 520-540 nm
Red: Ex = 570 nm, Em = 600-620 nm


<Experimental Conditons>
Instrument: Flow Cytometer
Filter: FITC (Ex = 488 nm, Em = 515-545 nm)

Application Data: Lipid Peroxidation Assay of Cumene hydroperoxide treated HepG2 Cells

HepG2 cells stained with this probe were stimulated with HBSS solution containing 200 µmol/l cumene hydroperoxide for 2 hours, and the fluorescence intensity was compared with control cells. The results showed a decrease in red fluorescence and an increase in green fluorescence in cumene hydroperoxide-treated cells compared to untreated cells, indicating that lipid peroxidation could be detected.

<Experimental Conditions>
Instrument: Fluorescent Microscope
Green: GFP filter (Ex = 450-490 nm, Em = 500-550 nm)
Red: TexasRed filter (Ex = 540-580 nm, Em = 600-660 nm)
Scale bar: 50 µm

Application Data: Lipid Peroxidation Assay of Erastin treated HepG2 Cells

HepG2 cells were stimulated with MEM medium containing 10 µmol/l Erastin for 2 hours and unstimulated cells were compared using this probe. The results showed a decrease in red fluorescence and an increase in green fluorescence in the stimulated cells compared to the untreated cells, indicating that lipid peroxidation could be detected.

<Experimental Conditions>
Instrument: Confocal Microscope
Green: Ex = 488 nm, Em = 510-550 nm
Red: Ex = 561 nm, Em = 600-630 nm)
Scale bar: 50 µm

References

Open References

 
No. Sample Type Instrument    Reference (Link)
1) Cell
(HTR-8/Svneo)
Microscope H. Yang, X. Zhang, Y. Ding, H. Xiong, S. Xiang, Y. Wang, H. Li, Z. Liu, J. He, Y. Tao, H. Yang, H. Qi, "Elabela: Negative Regulation of Ferroptosis in Trophoblasts via the Ferritinophagy Pathway Implicated in the Pathogenesis of Preeclampsia", cells2023, doi:10.3390/cells12010099.

 

Q & A

Q

How may samples can be tested by this kit?

A

Please refer to the  followings: 
・96-well plate: 2 plates
・ibidi 8-well plate: 12 plates
・35 mm dish: 10 dishes
・6-well plate: 10 well

 

Q

Is there a positive control experiment for lipid peroxidation assay?

A

The manual includes examples of detection using tert-butyl hydroperoxide (t-BHP) or cumene hydroperoxide-treated HepG2 cells. (Please refer to steps 1-5 in Application Data 1 and Application Data 2). 

Q

May I use buffers instead of medium to prepare the working solution?

A

It is not recommended to prepare the working solution with buffers other than medium. BDP 581/591 C11 is high fat solubility, prepared in a serum-free medium may cause the probe to deposit, please use a serum-containing medium for preparation.

Q

When using a flow cytometer in which step should I detach the cells from the plate?

A

Please follow step 6 of the manual, after the treatment and washing with HBSS, detach the cells with trypsin.

Q

Can I use other buffers in stead of HBSS to wash the cells after the staining?

A

Yes, you may use PBS, Hanks’ HEPES buffer, and a medium to wash the cells.

Handling and storage condition

Specification
Appearance: Purple Solid
Purity (HPLC): 90.0%
Handling and storage condition
Store in a cool, dark place.
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