storing membranes - (Aug/17/2007 )

i've probed my membrane with both antibodies and washed, but then realized that the developing machine is broken so left the membrane in 4C in PBS
how long can i store it like this? should i re-probe it before developing?

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signal will decrease over the time; the peroxidase activity if you use HRP-labeled sec Ab´s is rather instable; I think you have only some hours time (depending of course of the strongness of signal); are there no alternatives in neighbor labs?

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it may be better to warp the membrane and store it a 4oC, rather than keep it in PBS over a few days as the primary antibodies may disociate from the membrane.

Besides, the developing machine may take up to a week to fix.

When the developing machine is fix...
Just incubate the membrane with secondary antibodies for 1 hr or overnight at 4oC, wash membrane and add ECL, finally develop the film.

Hope this help.

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the best thing to have done is to complete the blot, expose the film and then hold the film to be processed when the machine is fixed or process it manually. this way you don't have to worry about the viability of the conjugated enzyme, stability of the antibody bond or anything else that someone may think of that may be a problem if you wait.

just remember to keep the film in the dark until processed.

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