For immunohistochemistry analyses cells were seeded on x-well Tissue Culture Chambers (Sarstedt, Nümbrecht, Germany) and transfected for 48 h. Cells were fixed with 4% formaldehyde for 15 min and permeabilized with 0.2% Tween20 for 5 min in phosphate-buffered saline. Cells were stained for 1 h with the primary antibodies: anti-MiD51 (1:100) (Proteintech), anti-Tom20 (1:100) (Abcam, Cambridge, UK), anti-insulin (1:100) (Abcam) and anti-LC3 (1:100) (Sigma). Cy5 or FITC-coupled secondary antibodies (1:250) were used for visualization (Molecular Probes Invitrogen, Darmstadt, Germany). Cells were mounted and counterstained using Roti®-Mount FluorCare DAPI (Roth, Karlsruhe, Germany) and analyzed using a Fluoview FV10i confocal microscope (Olympus, Hamburg, Germany).
Cy5 or fitc coupled secondary antibodies
Manufactured by Thermo Fisher Scientific
Sourced in Germany
Cy5 or FITC-coupled secondary antibodies are laboratory reagents used in immunodetection techniques. They consist of secondary antibodies that are conjugated to fluorescent dye molecules, either Cyanine 5 (Cy5) or Fluorescein Isothiocyanate (FITC). These antibodies are designed to bind to primary antibodies, allowing for the detection and visualization of target proteins or cellular components in various applications such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti insulin, by Abcam (2 mentions)
Fluoview fv10i confocal microscope, by Olympus (2 mentions)
Roti mount fluorcare dapi, by Carl Roth (1 mentions)
Anti tom20, by Abcam (1 mentions)
Anti mid51, by Proteintech (1 mentions)
Anti lc3, by Merck Group (1 mentions)
X well tissue culture chamber, by Sarstedt (1 mentions)
Anti fis1, by Santa Cruz Biotechnology (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
2 protocols using cy5 or fitc coupled secondary antibodies
1
Immunohistochemical Analysis of Mitochondrial Dynamics
2
Immunofluorescence Staining of FIS1 and Insulin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with 4% formaldehyde for 15 min and permeabilised with 0.2% Tween 20 for 5 min in phosphatebuffered saline. Finally, cells were stained for 1 h with the following primary antibodies: anti-FIS1 (1:100) (Santa-Cruz Biotechnology) and anti-insulin (1:100) (Abcam). Positive signals were visualised using Cy5 or FITCcoupled secondary antibodies (1:250) (Molecular Probes Invitrogen). The samples were counterstained with DAPI and mounted using VECTASHIELD mounting medium (Vector Laboratories, Peterborough, UK). Cells were analysed using a Fluoview FV10i confocal microscope (Olympus). A minimum of 10 sections per slide from three independent experiments were analysed using Fluoview-SW (Olympus) software.
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!