Immunofluoresence: Difference between revisions
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#Wash once with PBS | #Wash once with PBS | ||
#Permeabilize for 10 min with Triton X-100 (0.1% in PBS) | #Permeabilize for 10 min with Triton X-100 (0.1% in PBS) | ||
#Wash three times with PBS | #Wash three times with PBS (5 min each) | ||
#Block for 1-2h with 200 uL of BSA/Ovalbumin (1% of each in PBS) blocking solution | #Block for 1-2h with 200 uL of BSA/Ovalbumin (1% of each in PBS) blocking solution | ||
#Incubate overnight with primary antibody in blocking solution | #Incubate overnight with primary antibody in blocking solution | ||
#Wash coverslips 3 times 10 minutes with PBS | #Wash coverslips 3 times 10 minutes with PBS (5 min each) | ||
#Incubate in 500X secondary solution | #Incubate in 500X secondary solution | ||
#Wash coverslips 3 times 10 minutes with PBS | #Wash coverslips 3 times 10 minutes with PBS (5 min each) | ||
#Add 10 uL vectashield to glass slide and place cells on slide. Fix with nail polish | #Add 10 uL vectashield to glass slide and place cells on slide. Fix with nail polish | ||