Beas-2B cells (4.0 × 104 cells/well) were seeded in a 12-well plate which have been pre-placed cell climbing pieces at the bottoms, and were treated with indicated doses of NLD and THD for 18 h. Then, the cells climbing pieces were washed with PBS and fixed with −20 °C methanol/acetone (1:1) for 10 min. The cell climbing pieces were incubated with primary rabbit anti-Nrf2 antibody at 4 °C overnight. After 3 times washing in PBS, the cell climbing pieces were incubated with Alexa Flour 594 (Proteintech Group, IL, USA) for 1 h at room temperature in the darkness, and nuclei were covered with DAPI for 10 min. The fluorescence signals were imaged using Olympus BX53 fluorescence microscope coupled to Olympus DP73 digital camera (Tokyo, Japan).
Alexa flour 594
Manufactured by Proteintech
Sourced in United States
Alexa Fluor 594 is a red-fluorescent dye produced by Proteintech. It is a synthesized fluorescent label commonly used in biological research applications such as fluorescence microscopy and flow cytometry. The dye has an excitation maximum at 590 nm and an emission maximum at 617 nm.
Automatically generated - may contain errors
2 protocols using alexa flour 594
1
Nrf2 Protein Localization in Beas-2B Cells
2
Immunofluorescence Analysis of BV2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BV2 cells were seeded in 24-well plates at a density of 3*10 4 cells per well(500μL per well), incubated and treated as mentioned above. After xed with 4% paraformaldehyde for 20 min at room temperature, cell were permeabilized with 0.1% Triton X-100 for 20 min. Then blocking with 3% BSA for 1h at 37℃, and were incubated overnight at 4℃ with primary antibodies. (iNOS, Cell Signaling Technology , 1:400;NLRP3,1:50,Boste). On the owing day, After washed with PBS for three times, cells were incubated with secondary antibodies (Fluorescein(FITC)-conjugated A nipure Goat Anti-Rabbit IgG(H+L),1:400;Alexa Flour 594-conjugated A nipure Goat Anti-Rabbit IgG(H+L),1:400;Donkey Anti-Goat IgG H&L (Alexa Fluor® 555),1:200, Proteintech )in 1%BSA dilution for 1h at 37℃. The cells were treated with DAPI(Solabio) for 10 min. All images were captured with a uorescence microscope (Olympus) Results
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!