Revision 1

#95978

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
IF-IC, FC-FP

Product Usage Information

Supplied Reagent:
1. BODIPY 581/591 C11 (Lipid Peroxidation Sensor): Create a 10 mM stock solution by dissolving 1 mg of BODIPY 581/591 C11 (Lipid Peroxidation Sensor) into 198.26 μL of high-quality anhydrous DMSO.

Additional Reagents (Not Supplied):
1. Hanks' Balanced Salt Solution (HBSS)
2. DMSO (Dimethyl Sulfoxide), Sterile #12611

Protocol:
1. Incubate cells with 1 - 2 µM of BODIPY 581/591 C11 (Lipid Peroxidation Sensor) and 1 μM of Hoechst 33342 #4082 (optional) in cell culture media for 30 min.
2. Wash two times with HBSS.
3. Add desired treatments in HBSS and incubate as needed.
4. Examine cells by fluorescence microscopy. Refer to the Fluorescent Properties section for excitation and emission spectra.
5. For analysis by flow cytometry, dissociate adherent cells or collect suspension cells. Viability dye can be used if desired. Analyze on a flow cytometer at desired cell density.

Storage

Store lyophilized at -20°C. In lyophilized form, the product is stable for 12 months.

Description

BODIPY 581/591 C11 (Lipid Peroxidation Sensor) is a fluorescent dye that localizes to membranes and is used to measure lipid peroxidation in live cells. It is a highly sensitive fluorescent probe that has a shift in fluorescent emission upon oxidation of the polyunsaturated butadienyl portion of its fatty acid analog in live cells. When this occurs, fluorescence emission peaks shift from red (~590 nm) to green (~510 nm). BODIPY 581/591 C11 (Lipid Peroxidation Sensor) measures lipid peroxidation through the ratiometric shift in fluorescence between reduced and oxidized states of the probe.

Lipid peroxidation is the free radical oxidative degradation of lipids from excessive reactive oxygen species production. Oxidative damage to lipids produces lipid peroxides that can cause damage to cell membranes, leading to changes downstream in signaling pathways, eventually leading to cell death. Lipid peroxidation can occur during apoptotic cell death but has been recognized as a defining characteristic of the non-apoptotic cell death pathway ferroptosis. Ferroptosis is defined by iron-dependent, lipid peroxidation. An increase in ferroptosis occurs during aging, and pathologies including cancer, atherosclerosis, neurodegeneration, and cardiovascular disease.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

CST Logo
Confocal analysis of HT-1080 cells, untreated (left), treated with cumene hydroperoxide (200 μM, 2 hr; middle), or treated with cumene hydroperoxide (200 μM, 2 hr) after inhibiting with α-tocopherol (25 μM, 20 hr; right) using BODIPY 581/591 C11 (Lipid Peroxidation Sensor) (green and red) and Hoechst 33342 #4082 (blue).
Immunofluorescence Image 1: BODIPY 581/591 C11 (Lipid Peroxidation Sensor)
Confocal analysis of HT-1080 cells, untreated (left) or treated with erastin (10 μM, 22 hr; right), using BODIPY 581/591 C11 (Lipid Peroxidation Sensor) (green and red) and Hoechst 33342 #4082 (blue).
Immunofluorescence Image 2: BODIPY 581/591 C11 (Lipid Peroxidation Sensor)
Fluorescence analysis of HT-1080 cells incubated with 2 μM BODIPY 581/591 C11 (Lipid Peroxidation Sensor). Increasing concentrations of cumene hydroperoxide cause an increase in lipid peroxidation which is detected as increasing 488 MFI (green) and decreasing 564 MFI (red). MFI data were captured using an Operetta CLS high content analysis system.
Immunofluorescence Image 3: BODIPY 581/591 C11 (Lipid Peroxidation Sensor)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

CST Logo
Fluorescence analysis of HT-1080 cells incubated with 1 μM BODIPY 581/591 C11 (Lipid Peroxidation Sensor). Increasing concentrations of tert-Butyl hydroperoxide cause an increase in lipid peroxidation which is detected as increasing 488 MFI (green) and decreasing 564 MFI (red). MFI data were captured using an Operetta CLS high content analysis system.
Immunofluorescence Image 4: BODIPY 581/591 C11 (Lipid Peroxidation Sensor)
Flow cytometric analysis of live HL-60 cells, untreated (blue) or treated with tert-Butyl hydroperoxide (200 μM, 2 hr; green), labeled with BODIPY 581/591 C11 (Lipid Peroxidation Sensor) (2 μM, 30 min) (solid lines) or unlabeled (dashed lines). Data collected using blue laser excitation and FL1 emission.
Flow Cytometry Image 1: BODIPY 581/591 C11 (Lipid Peroxidation Sensor)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.