you donot need to use agar block for IHC or ICC, whatever you want to do.
Kindly follow the protocol.
Fix the cells with 4% formaldehyde for 20 minutes at room temperature by adding formaldehyde directly to the culture media (see page 5) and adjust to approximately 1 x 106 cells/mL.
2. Add 1 mL of the cell solution to a 1.5 mL microfuge tube.
3. Spin down the cells for 30 seconds in a microfuge.
4. Pour off the supernatant and resuspend the cell pellet in 1 mL of deionized H2O.
5. Spin down the cells for 30 seconds in the microfuge.
6. Pour off the supernatant and resuspend the cell pellet in 200 mL of deionized H2O.
7. Add 5 mL of the cell suspension to a gelatin-coated slide (3 spots per slide) and smear with the side of a pipette tip.
8. Place the slide on a hot plate (low heat setting) and allow the liquid to evaporate.
9. Check the slide under a microscope and make sure that there are no salt crystals.
If salt crystals are observed, wash the slide with deionized H2O.
10. Surround the cell spot with a hydrophobic barrier using a barrier pen and air dry.
11. The slides can be stored at 2–8 °C for up to 3 months or they may be stained immediately.
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