incorrect blocking in immunocyto.... - (May/06/2010 )

Hi,

Had a little hiccup with my immuno by mistake I blocked my cells with goat serum and one of my antibodies is raised in goat.... so just wondering does anyone know can cells be re-blocked or can the goat serum be reomved? or is there no hope-should I chuck the cells? They take ages to culture so even though it's only one prep it's a hassel...
Anyone with any ideas would be great,
thanks!!

if your secondary antibody is made in goat then goat serum is a good blocking agent.

Hi,

No it's my primary that is raised in goat, so my secondary is anti goat, so I'm trying to see if I can re-block with horse or something to still be able to probe for my primary without all the non specific binding to the goat serum....

then, unless you have another primary you probably won't be able to use the cells. they are blocked. i know of no way to remove the block without damaging the cells.

mdfenko on May 7 2010, 09:36 PM said:

i second that.. i have never come across such a thing... although if u happen to stumble upon sumthing useful.. please do share it with us!!!
Best luck!!

antibody is made in goat then goat serum is a good blocking agent.
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anokhi on May 16 2010, 01:27 PM said:

not if it is the primary antibody that is made in goat.

Well, I tried many washes with Triton X and re-blocking... but nope , the cells were no good! so chucked them! Wont be making that mistake again!

mdfenko on May 18 2010, 10:29 AM said: