2 replies to this topic

[philman]

my protocol I have been given says to use the antibody against my surface protein of interest at a final concentration of 10ug/ml in 1ml of media for live cell staining.

Given that the antibody is supplied in a vial labelled 100ug total, this seems an awful lot of antibody for 1 slide, am I missing something here in my calculations or do I really have to use 1 vial per 10 slides?

[philman]

ok this is probably in the wrong section as I am doing imaging of cells in culture not tissue, ill re-post it in cell biology or somethng

[casandra]

philman, on 06 April 2011 - 05:20 AM, said:

my protocol I have been given says to use the antibody against my surface protein of interest at a final concentration of 10ug/ml in 1ml of media for live cell staining.

Given that the antibody is supplied in a vial labelled 100ug total, this seems an awful lot of antibody for 1 slide, am I missing something here in my calculations or do I really have to use 1 vial per 10 slides?

I've not done live cell staining but are you gonna flood the whole slide with 1 ml? Usually, we draw beautiful tiny circles (2 to 3) per slide with those pricey immunocyt pens and then just incubate with 80-100 ul of antibody solution...or you grow your cells in these special slides with chambers.... but I'm not sure if this is how you do your staining...and usually for immunocytochemistry, we use more concentrated  ab solutions....