Native Gel Staining?? - So I can electrolute proteins by cutting gel slice (Jun/14/2005 )

Thought I would poke my head into this board as well.


I need to stain my native PAGE gels for my GST fusions proteins. However I need to cut out the gel slices and electrolute the proteins w/out any stain bound to proteins so I can use the proteins later in downstream pro/pro interactions experiments.

I would be appreciative of any help.

Thanks

---

Viper,

To cut proteins slices from PAGE without stainning the gel, you can do this. Run two PAGE simultanily (with same sample, sample separation conditions sample) and stain one gel. With this, you can see where are your band of interest. Just cut on the unstained gel at the same place where your protein are located on the stained gel.

Hope that it will help you

---

Actually I know that...however I can't do it because the variability in migration is just enough that I could slice into a degradation product below teh full length product..

thanks anyway

---

May be you can do westernblotting and visualize the protein by any of the destainable stain.

---

Hi viper

There is reversed (it stains the gel not the protein) and reversible (you can destain easily) staining KIT. You can stain your gel, cut the bands you wish out and destain it.

I have been using it conveniently (I stain to check the band, take a picture and destain the gel and transfer to membrane for western blot).

Unfortunately, the KIT I am using is Japanese made and I do not know Japanese to get you more information!

---