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Author Topic: Antigen Blocking  (Read 5933 times)

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Offline Amanita-Virosa

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Antigen Blocking
« on: April 20, 2006, 07:15:49 AM »
Hi,

I've recently being trying to establish whether my antibody is binding specifically to its target or not.  In order to test this I have incubated the antibody with its antigen at various concentrations before performing ICC in an attempt to abolish staining of my cells.

I have however had some odd results that I cannot explain; At the higher antigen concentrations instead if staining being decreased or absent it acutally seems to increase - sometimes quite dramatically.  Does anyone have any suggestions as to what may be causing this. I am very confused!!   :?

Thanks.

Antigen Blocking
« on: April 20, 2006, 07:15:49 AM »

Offline richard03

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Antigen Blocking
« Reply #1 on: April 22, 2006, 02:30:35 PM »
The antigen may bind to tissue unspecifically. Any details about your method of blocking may help.

Offline ImmunoNYC

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Re: Antigen Blocking
« Reply #2 on: April 24, 2006, 12:16:07 AM »
We need more details to know what is happening here.[/color]

Quote from: "Amanita-Virosa"
Hi,

I've recently being trying to establish whether my antibody is binding specifically to its target or not.  In order to test this I have incubated the antibody with its antigen at various concentrations before performing ICC in an attempt to abolish staining of my cells.

I have however had some odd results that I cannot explain; At the higher antigen concentrations instead if staining being decreased or absent it acutally seems to increase - sometimes quite dramatically.  Does anyone have any suggestions as to what may be causing this. I am very confused!!   :?

Thanks.

Re: Antigen Blocking
« Reply #2 on: April 24, 2006, 12:16:07 AM »