u/bluemooninvestor

Hello everyone,

I have been using Pierce Desalting Spin columns for peptide cleanup, and it required 300 uL of 50% ACN to elute.

Now, I have to use the Pierce C18 columns. Despite both of them being small spin columns, the C18 column protocol says that only 20 uL of 70% ACN should be used for elution.

My question is, isn't 20 uL a very small volume? Why is there such a huge difference in elution volume between the two spin columns (Desalting v C18 one)? Lastly, is there any general understanding in the proteomics community to use a larger elution volume with the Pierce C18 spin columns.

The protocol also says one may use 70% ACN with 0.1% TFA, but no idea why that is not the standard elution solution for this setup.

I understand that most hardcore proteomics labs would probably not be using Pierce C18 spin columns, but this is what I am using as a part-time proteomics guy. I don't have any other C18 setup, and I will send the cleaned-samples to a mass-spec facility.

And advice on the Pierce C18 spin columns is greatly appreciated.

Recently I have run into Lip-ms, and there are quite a few high impact papers describing the technique. But there are really few papers who have actually used it for doing science.

I think it is the same for several other techniques like proximity-labelling MS, certain advanced variants of thermal proteome profiling etc.

Why do you think it is like this? If these techniques are so great, why aren't they being used for actual science on a much greater scale? Or is my assumption wrong?

Excited to listen to your views.

For perspective, I am a cancer biologist trying to do omics.

I have submitted a paper to JPR for the first time. It has been three weeks and the status is still "Editorial Review".

Does the manuscript status change to Peer Review or something like that when it's under actual review? Or does it mean it's still with the editor only?