no reference in the westerns blot (like actin) - are these results publishable? (May/12/2006 )
as you might already know im writting up some paper of results which are not mine....so yesterday while writting up the result section we have discovered that my freind (who done the experiments and is not here anymore) didnt include any control in her westerns....like actin you know....to have some stable reference....i just cant repeat all her experiments...and i dont think she left the membrane so i can reprobe with actin or something...is there anything i can do? are the results publishable without any reference in the westerns? thanx for any input...
well it depends, where do you want to publish it? if It is "Nature Biotechnology" forget the result and repeat it.
i still didnt think about which journal....do you have any advice??
I think in general it`s easier to write a paper if you have a certain journal in mind, as you have to format your paper in the journal-way.
You might save a lot of time by considering the journals other publications and see how results etc. are prepared and presented.
Regarding actin and western blot things: it depends on your experiments and the results you want to show. If it is a knock down or something like this you would need actin defintely. If it`s just to proof that anything is expressd etc. you might not need it. Sometimes it is accepted to declare that for example Ponceau-Red staining was performed and the amount was equal, or you explain the you load the SDS-Page after Bradford-measurement. This might be acceptd, though this is not really convincing I´m afraid... . Anyway I have seen this in some papers.
Good luck for your manuscript.
bomber, thanx a lot for your help! she has done westerns to show an increase in the band intensity... ...so in this case im afraid control is very important right?
I would say yes.
What you could do is to choose a formulation like Ponceau-Red staining showed equal intensities; but if I figure it right you haven't seen any Ponceau -Red or Bradford assays etc...., so I wouldn't recommend it becuase you are responsible for the things you write in the paper.
Try to repeat an experiment. If you still have the lysates it shouldn't be a time consuming work to do a western...right?
i suppose you have used the same cell number in all your wells...
ponceau staining is not sufficent for an intern reference.
Best is a control antibody, and i'm afraid that you'll have to repeat all the exps...