Anti Lamp-1 or Lamp-2 for COS cells? - (Nov/04/2005 )

Hi, I'm looking for lysosome markers (ab anti-Lamp1 or anti-Lamp-2 if it's possible) in COS cells but all commercially available ab are for human cells. Do you know if it's usual that antibodies anti-human cross-reacts with monkey antigens, or if there is any good antibody anti-lamp1 in monkey?

Lysotracker red as lysosome marker didn't work at all.

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Lysotracker Red from Molecular Probes works, use it 10 fold more concentrated for 30 minutes or more at 37 degrees and it will stain. Not sure if it was suppose to work at the manufacturer's 50 nM concentration or not but i used 500-750 nM and it worked fine. Let me know and i'll give you more details.

Now, i myself am looking for a specific antibody that works for lysosomes in COS. Do you know of a commercially available one that works and stains lysosomes in COS cells?

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Did you possibly permeabilize your cells for analysis? Becasue the lysotracker will leak out then.
As for the antibodies, I would try the anti human antibodies unless it has been specifically explained that they do not work in monkey. Human and monkey proteins are very similar, even the DNA sequences are so similar that most siRNAs against human genes can be used in monkey derived cell lines.
I hope it helps.
Cheers,
Diril

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That assumption was my assumption until i tested those antibodies. I have not found a human lamp-1 antibody to work on monkey cells. Anyway, you do not need to permeabilize the cells prior Lysotracker treatment. It is a basic amine fluorophore that you can use directly with your cells. If anyone has found an anitibody that works, please let me know!

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