October 18, 2021, 01:06:04 PM
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Topic: x ray development  (Read 2450 times)

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Offline shininglight

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x ray development
« on: August 14, 2007, 07:06:59 AM »
hi, could someone explain how after a western blot is treated with substrate it is developed in dark room with x ray film and where the antibody attached to protein on the membrane a black spot appears , i know the procedure i just dont understand how it happens, thanks

Offline sdekivit

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Re: x ray development
« Reply #1 on: August 14, 2007, 02:03:41 PM »
after transferring the proteins to the sheet antibodies (primary antibody) is incubated that specifically binds to the protein pof interest.

next a second antibody directed against the constant region of the primary antibody (say the first antibody is derived from a mouse, the second antibody derived from a goat is directed to mouse antibodies)

--> the second antibody is conjugated with a radioactive label that can be made visible on an autoradiogram (x-ray film)

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