Protocol- Anti-FITC Immuno Template

Schedule for FITC Immuno Round Completion:

Single labeling:
Friday- Sections into Primary (5x5min rinses, 30min pre-incubation/pre-absorption)

Monday- FITC(DK)anti(RB/MS) 1:500 in LTBS (5x5min rinses, 30min pre-incubation/pre-absorbtion,  
             2-3hrs in 2˚
             Alexa488(RB)anti(FITC) 1:500-2k? in TBS ~36hrs

Wednesday- Cy2(MS)anti(FITC) (1:1-2k) in TBS ~2-3hrs

Double labeling: 
Friday- Sections into Primary (5x5min rinses, 30min pre-incubation/pre-absorption)

Monday- Biot(DK)anti(RB) 1:500 in LTBS (5x5min rinses, 30min pre-incubation/pre-absorbtion, 2hr)
            FITC(DK)anti(MS) 1:500 in LTBS (5x5min rinses, 30min pre-incubation/pre-absorbtion, 2hr)
            Cy2(MS)anti(FITC) 1:500-2k in TBS ~36hrs

Wednesday- Cy3SAv 1:500 in TBS ~2hrs
                 

OR

Friday- Sections into Primary (5x5min rinses, 30min pre-incubation/pre-absorption)

Monday- Biot(DK)anti(MS) 1:500 in LTBS (5x5min rinses, 30min pre-incubation/pre-absorbtion, 2hr)                   
               FITC(DK)anti(RB) 1:500 in LTBS (5x5min rinses, 30min pre-incubation/pre-absorbtion, 2hr)
               Alexa 488(RB)anti(FITC) 1:500 in TBS ~36hr

Wednesday- Cy3SAv 1:500 ~2hrs
               (5x5min rinses)