1999 FC Control Files
Microbiology 542 Immunology Lab
Splenocytes:
- Autofluorescence
Use these values for red/FL2 blank.
- Second antibody only
Use these values for green/FL1 blank.
- PE-CD4
Thymocytes:
- Autofluorescence
Use these values for red/FL2 blank.
- Second antibody only
Use these values for green/FL1 blank.
- Thy-1 (indirect, FITC)
- PE-CD4
There are no controls for unwanted binding of isotype-matched
first layer antibody in indirect staining. I did one for IgG2b
(see M1/69 in Antibodies Available) but the resulting intensities
were slightly below those for autofluorescence. Therefore we'll
assume that there is no unwanted binding of first layer antibody.
There are no controls for unwanted binding of PE-conjugated
or FITC-conjuagated
antibodies in direct staining (such antibodies are too expensive).
Therefore we'll assume that such unwanted binding is insignificant.