Mike Barnkob:Protocols/Visualization/Immunofluorescence on cell lines: Difference between revisions
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'''Prepare:''' | '''Prepare:''' | ||
* Fixative: 1 ml 4% formaldehyde | * Fixative: 1 ml 4% formaldehyde <br />'''Note:''' 250 ul 16% formaldehyde in 750 ul PBS. | ||
* Blocking solution: 1% bovine serum albumin (BSA) in PBS | * Blocking solution: 1% bovine serum albumin (BSA) in PBS <br />'''Note:''' 1g BSA to 100ml of PBS. <br />'''Note: BSA is in common reagens fridge. | ||
===Setup and controls=== | ===Setup and controls=== | ||
Revision as of 02:27, 27 May 2015
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
Note: 250 ul 16% formaldehyde in 750 ul PBS. - Blocking solution: 1% bovine serum albumin (BSA) in PBS
Note: 1g BSA to 100ml of PBS.
Note: BSA is in common reagens fridge.
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.
