I have tried 1% BSA in PBS and it work slightly better than my previous results but did not obtain the clarity that I need. So please suggest me any appropriate techniques to remove the background noise completely as posiible.
hi,
-At this stage, you can take an attempt by increasing the concentration of BSA to 3% or 5%. It will help to reduce the background.
-Sometimes, poor fixation and permeabilization step also causes background noise.
-Antibody concentrations and incubation times cannot be ignored for such noise because antibody binding within the incubation period should be optimum to get the best fluorescence intensity, so the background could be nullified.
-Then, a Secondary antibody must be standardized to achieve the result.
-A dark environment, fresh reagents without any contamination should be used to avoid any unwanted things.
Regards
Saurabh
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