Mike Barnkob:Protocols/Visualization/Immunofluorescence on cell lines
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Immunofluorescence protocol
This is a protocol for doing IF (immunofluorescence) on adherent and non-adherent cell lines.
This protocol was last updated on 27th of May 2015.
This protocol is not yet complete.
Reagents
- Coverslips: round Ø 13 mm for adherent cells (VWR, cat. 631-0141) or 24x60 mm for non-adherent cells (VWR, cat. 631-0150).
- Microscopy slides (VRW, cat. 631-0909).
- PBS
- Formaldehyde, 16%
Prepare:
- Fixative: 1 ml 4% formaldehyde (250 ul 16% formaldehyde in 750 ul PBS).
- Blocking solution: 1% bovine serum albumin (BSA) in PBS (1g BSA to 100ml of PBS).
Setup and controls
- Negative control: one samples prepared as others, but without antibody staining.
- Isotype control: one samples prepared as other, but has been stained with an irrelevant antibody of the same isotype which has no specificity to the protein of interest.
- Secondary control: : one samples prepared as other, but stained only with the secondary antibody.
