After appropriate treatment, cells were fixed with 4% paraformaldehyde for 15 min, permeabilised with PBS/0.2% Triton X-100 (Sigma-Aldrich) and blocked with PBS containing 10% FBS or 10% goat’s serum (Gibco). For protein detection, primary antibodies were incubated in PBS/1% FBS or goat’s serum overnight at 4°C. Cells were then incubated for 1 h at RT with conjugated secondary antibodies in PBS/10% FBS or goat’s serum. Where appropriate, cells were counterstained with 4′,6′-diamidino-2-phenylindole (DAPI). Images were captured using Carl Zeiss LSM880 confocal microscope (Jena, Germany) and analysed using Image J software (National Institutes of Health, Bethesda, MD, USA).
Goat s serum
Manufactured by Thermo Fisher Scientific
Sourced in United States
Goat's serum is a biological fluid derived from the blood of goats. It is a complex mixture of proteins, lipids, and other biomolecules that can be used as a supplement in various cell culture and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using goat s serum
1
Immunofluorescence Staining of Cells
2
Immunofluorescence and Live-Cell Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded on 21-mm borosilicate glass cover slips, 8 chamber polystyrene vessel CultureSlides (Falcon, Fisher Scientific, Loughborough, UK) or 35mm imaging dishes (Ibidi, Thistle Scientific, Uddingston, UK). Images were captured using Carl Zeiss LSM880 confocal microscope (Jena, Germany) and images were analysed using Image J software 18 (National Institutes of Health).
For fixed cell imaging: Cells were fixed with 4% paraformaldehyde (PFA) for 15 min, permeabilized with PBS/0.2% Triton X-100 (Sigma Aldrich) and blocked with PBS containing 10% goat's serum (Gibco) and 2.5% Human TruStain FcX (BioLegend, San Diego, USA). Primary antibodies (Table S2) were incubated overnight at 4°C and conjugated secondary antibodies for 1hr at RT. Where appropriate, cells were counterstained with 4′,6′-diamidino-2phenylindole (DAPI) or mounted with Vectashield mounting medium for fluorescence with DAPI (Vector Laboratories, Peterborough, UK).
For live cell imaging: Cells were grown in 35mm imaging dishes (Ibidi) and maintained at 37 o C and 5% CO2 in live-cell imaging chamber attached to Carl Zeiss LSM880 confocal microscope.
Images were captured every 2 minutes at x40 magnification over a 12hr time period.
For fixed cell imaging: Cells were fixed with 4% paraformaldehyde (PFA) for 15 min, permeabilized with PBS/0.2% Triton X-100 (Sigma Aldrich) and blocked with PBS containing 10% goat's serum (Gibco) and 2.5% Human TruStain FcX (BioLegend, San Diego, USA). Primary antibodies (Table S2) were incubated overnight at 4°C and conjugated secondary antibodies for 1hr at RT. Where appropriate, cells were counterstained with 4′,6′-diamidino-2phenylindole (DAPI) or mounted with Vectashield mounting medium for fluorescence with DAPI (Vector Laboratories, Peterborough, UK).
For live cell imaging: Cells were grown in 35mm imaging dishes (Ibidi) and maintained at 37 o C and 5% CO2 in live-cell imaging chamber attached to Carl Zeiss LSM880 confocal microscope.
Images were captured every 2 minutes at x40 magnification over a 12hr time period.
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!