Cells were fixed 4% paraformaldehyde in PBS for 20 min, permeabilized with 0.1% Triton X-100 in PBS for 5 min, then washed in PBS, then in PBS supplemented with 1% BSA and 1% rabbit IgG blocking reagent (Thermo Fisher Scientific) for 5 min, then in PBS. Anti-clathrin (ab21679; Abcam) and anti-EGF receptor (D38B1; Cell Signaling) were labeled with Alexa488- and Alexa647-Zenon rabbit IgG labeling kits, respectively (Thermo Fisher Scientific), according to the manufacturer’s instructions, and cells were then incubated with both antibodies (both diluted 1:100 in PBS–1% BSA) for 20 min. After washing thrice in PBS, imaging was performed in PBS instead of mounting medium to avoid squashing the cells (and potentially biasing the clathrin or EGFR micropattern colocalization).
Rabbit igg blocking reagent
Manufactured by Thermo Fisher Scientific
1% rabbit IgG blocking reagent is a laboratory product designed to block non-specific binding of rabbit immunoglobulin G (IgG) in various immunoassay and immunohistochemistry applications. It is a pre-formulated solution that can be used to reduce background signal and improve the specificity of rabbit IgG-based detection systems.
Automatically generated - may contain errors
2 protocols using rabbit igg blocking reagent
1
Visualizing Clathrin and EGFR Colocalization
2
Immunofluorescence Imaging of Clathrin and EGFR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed 4 % paraformaldehyde in PBS for 20 min, permeabilized with 0.1 % Triton X-100 in PBS for 5 min, then washed in PBS, then in PBS supplemented with 1% BSA and 1% rabbit IgG blocking reagent (ThermoFisher) for 5 min, then in PBS. Anti-Clathrin (Abcam, ab21679) and anti-EGF Receptor (Cell Signaling, D38B1) were labelled with Alexa Fluor 488-and Alexa Fluor 647-Zenon Rabbit IgG labelling kits respectively (ThermoFisher), according to the manufacturer's instructions, and cells were then incubated with both antibodies (both diluted 1:100 in PBS-1% BSA) for 20 minutes. After washing thrice in PBS, imaging was performed in PBS instead of mounting medium to avoid squashing the cells (and potentially biasing the Clathrin or EGFR / pattern colocalization).
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!