3 replies to this topic

[kaveh]

I am preparing my figures for a manuscript. For a western blot, I have probed the blot with two different antibodies. Those blots are in different figures of the same manuscript. Since the loading control for both of them are the same (I have re-probed the same membrane with actin), can I use that loading control in two separate figures?

Figure.1
Ab#1 + actin

Figure.2
Ab#2 + actin

I appreciate your inputs.

[casandra]

kaveh, on 20 May 2012 - 08:48 PM, said:

I am preparing my figures for a manuscript. For a western blot, I have probed the blot with two different antibodies. Those blots are in different figures of the same manuscript. Since the loading control for both of them are the same (I have re-probed the same membrane with actin), can I use that loading control in two separate figures?

Figure.1
Ab#1 + actin

Figure.2
Ab#2 + actin

I appreciate your inputs.

You haven't attached any figures. But why use the same loading control for both blots? Bec one of them sucked (bad blot or the loading was not equal?).

[kaveh]

casandra, on 21 May 2012 - 05:04 AM, said:

kaveh, on 20 May 2012 - 08:48 PM, said:

I am preparing my figures for a manuscript. For a western blot, I have probed the blot with two different antibodies. Those blots are in different figures of the same manuscript. Since the loading control for both of them are the same (I have re-probed the same membrane with actin), can I use that loading control in two separate figures?

Figure.1
Ab#1 + actin

Figure.2
Ab#2 + actin

I appreciate your inputs.

You haven't attached any figures. But why use the same loading control for both blots? Bec one of them sucked (bad blot or the loading was not equal?).

I probably didn't explain it well. Here is the question:
Suppose that I want to look at protein X in two different samples (control vs. experiment) when I am done with protein X, I re-probe the same membrane to look at protein Y. And finally, I re-probe the very same membrane with actin as a loading control. Obviously, because protein X and Y are on the same blot, the same actin would be a loading control for both them, am I wrong?

Now my question is whether it's ok to use the same actin picture for protein X and Y, while they are on separate figures.

[doxorubicin]

It would be a non-standard thing to do.  Just make it a single figure.