Cell fixation DTAF

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Cell fixation DTAF

Postby elflick » Jan 13 2012 3:23 pm

I want to incubate cells with 5-DTAF labelled pectins. These will be DLD1 and HCT116 colon cancer cells, and also RAW murine macrophages.
I realise I will have to fix the cells in paraformaldehyde afterwards, then put them on a slide for visualising under a fluroscence microscope.
I've never done anything like this before and have not yet found anyone who knows - so please bear with me if the questions seem stupid!

I was wondering how long the slides would keep for? e.g would they keep for a week before I look at them?

Also, after fixing the cells, I wanted to add an antibody for a protein (galectin-3, mouse), and then a fluorescently labelled secondary antibody, so I could then visualise whether the protein and the pectins are associating with eachother.

Again, how long would the slides keep for?

I'm just starting to research how to do all this, so if anyone has any tips or websites I could look up that would be great.
Do you know what kind of labelled secondary-antibody I should use that would be a different colour to DTAF?
Many thanks for any help you can give!
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