Cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and incubated with blocking buffer samples, and then with a rabbit monoclonal antibody specific to vimentin (1:100 dilution, CST) and mouse monoclonal antibody specific to E-cadherin (1:50 dilution, CST) at 4 °C overnight. After extensive wash, secondary anti-rabbit IgG conjugated with Alexa 647(CST) or anti-mouse IgG conjugated with Alexa 594 (Proteintech) were incubated. The samples were then fixed and analyzed via confocal microscopy. Cell fluorescence intensity was quantified with Image J.
Alexa 594
Manufactured by Proteintech
Alexa Fluor 594 is a red-fluorescent dye used for labeling proteins, nucleic acids, and other biomolecules. It has an excitation maximum of 590 nm and an emission maximum of 617 nm. Alexa Fluor 594 is a commonly used fluorescent probe in various applications, including immunohistochemistry, flow cytometry, and fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alexa 594
1
Immunofluorescence Analysis of Vimentin and E-Cadherin
2
DNA Damage Assessment Using γH2AX and 53BP1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA damage was determined using γH2AX and 53BP1 as readouts. After UV irradiation or treatment with DNA damaging agents, cells were allowed to recover for 1 h at 37°C before fixation with 4% formaldehyde in PBS for 20 min at room temperature. The fixed cells were washed twice with PBS and permeabilized in 0.2% Triton X-100 in PBS for 5 min. After washing twice with PBS, the slides were blocked with blocking buffer (0.2% Triton X-100, 5% BSA in PBS) for 60 min and incubated with anti-γH2AX (EMD Millipore) or anti-53BP1 (Abcam) antibody. After washing with PBS, the slide was incubated with the anti-rabbit secondary antibody Alexa 594 (Proteintech). The slides were visualized using Leica DM400 and quantified by ImagePro.
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!