ImmunostainingEmbryos.md

Immunostaining Embryos

Reagents

*	4% PFA in 1X PBS
*	0.5% Triton-X in 1X PBS
*	1X PBS
*	1% BSA in 1X PBS
*	Antibodies

Protocol made for a single slide. Adjust measurements for your needs.

*	Slide is kept in a humid chamber throughout the experiment

1. Take a mini pap pen and trace a rectangle around your sample. Let dry
2. Add 200uL of fixation solution(4% PFA in 1X PBS) to cover the cells for 10 minutes
3. Incubate fixed cell with Permeabilization solution (0.5% Triton-X in PBS) for 10 minutes
4. Add 200 uL of 1% BSA in 1X PBS at room temperature. Leave for 30mins - 1hr
5. Add 200uL of primary antiboy in blocking solution for 1-2hrs in the 4C fridge
6. Wash 3X in 1X PBS
7. Add 100uL of secondary antibody in blocking solution for 1-2hrs at room temperature
8. Wash 3X in PBS
9. Mount the seal chamber with diamond shield. Can be imaged right away or stored at 4C