A375 cells were treated with LPS (1 µg/mL) for 24 h and then fixed in 4% paraformaldehyde (PFA). The fixed cells were permeabilized using 100% methanol and stained with an anti-STAT3 antibody (CST: #12640; MA. USA) overnight at 4 °C. Cells were then incubated with corresponding secondary antibodies at room temperature for 1 h. Finally, the slices were mounted with DAPI-containing mounting medium (Abcam) and imaged using a fluorescence microscope (Leica DMI3000 B).
Dapi containing mounting medium
Manufactured by Abcam
Sourced in China
DAPI-containing mounting medium is a liquid solution designed to mount and preserve biological samples for microscopic analysis. The medium contains the fluorescent dye DAPI, which binds to DNA and produces a blue fluorescent signal when excited by ultraviolet light. This allows for the visualization and identification of cell nuclei within the mounted sample.
Automatically generated - may contain errors
Lab products found in correlation
Dmi3000b, by Leica (1 mentions)
Snap cell tmr star, by New England Biolabs (1 mentions)
Ab104139, by Abcam (1 mentions)
X tremegene hp reagent, by Roche (1 mentions)
Alexa 488 green, by Cell Signaling Technology (1 mentions)
Sp5 laser scanning confocal microscope, by Leica (1 mentions)
Alexa 555 red, by Cell Signaling Technology (1 mentions)
3 protocols using dapi containing mounting medium
1
STAT3 Activation in A375 Cells
2
Subcellular Localization of Endogenous and Exogenous BAP1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For detecting endogenous BAP1, parental, TNPO1-, KPNA1-, and KPNA2-KO lines were fixed with 4% formalin, stained with mouse anti-BAP1 antibody (1:100, clone #C-4, sc-28383; Santa Cruz Biotechnology), and anti-mouse Alexa Fluor 594 antibody (1:2,000, A21203; Invitrogen), and mounted in DAPI-containing mounting medium (AB104139; Abcam). For detecting exogenous BAP1, HEK293T cells were transfected with SNAP-tagged BAP1 using X-tremeGENE HP reagent (Roche) for 2 d, and SNAP-tagged BAP1s were live-labeled with SNAP-Cell TMR-star (New England BioLabs) overnight. Labeled cells were fixed with 4% formalin and mounted in DAPI-containing mounting medium (Abcam). Immunofluorescence images were taken using a Leica DMI6000 microscope with an HCX PL FL 63×/1.4-NA objective lens and an Andor Luca R EMCCD sensor. Image acquisition was performed with the application of MetaMorph Multi-Wavelength acquisition software (Molecular Devices). Phenotypic distribution of BAP1 was manually counted in three independent assays, with ≥250 cells counted in each experiment.
3
Immunofluorescence Staining of Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For IF staining, cells were grown on glass coverslips and then fixed with 4% paraformaldehyde for 15 min at room temperature. Following fixation, the cells were permeabilized using 0.5% Triton X-100 for 15 min and blocked with 5% goat serum for 1 h. Then, the cells were incubated with primary antibodies overnight at 4 °C and appropriate secondary antibodies conjugated with Alexa 555 (red) or Alexa 488 (green) (Cell Signaling Technology). DAPI-containing mounting medium (Abcam, Shanghai, China) was used to stain nuclei and mount cells. A Leica SP5 Laser Scanning Confocal Microscope (Leica Microsystems, Buffalo Grove, IL, USA) was used to capture and analyze images.
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!