Juan José Nieto Fontarigo

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Questions related from Juan José Nieto Fontarigo

Can I do western blot in human serum? Should I charge by volume or by protein quantity?

We want to do western blot in human serum in order to validate some proteins with changes resulting from iTRAQ-MS/MS data. Is it possible to do western blot directly in human serum without...

23 April 2019 7,806 4 View

Which is the best protocol for plasma exosomes purification?

We did ultracentrifugation to purify exosomes, but we only obtain 4 ug of protein (BCA assay of intact exosomes) from 2 mL of plasma. This is a very low yield. We need at least 40 ug to do flow...

05 January 2018 6,962 3 View

Why my IEF doesn´t reach enough voltage?

I am trying to do an 2-DE after ACN precipitation but when I do the IEF, I do not enough reach the voltage that I need (for example, in the first step, I need 250V but I only have 15V and the...

09 March 2017 4,578 4 View

How can I resolubilize proteins after acetone precipitation?

They were resuspended in 10 mM phosphate buffer but were not resolubilized

25 November 2016 1,725 4 View

Why if I load 30 ug of low molecular weight proteins in a SDS-PAGE gel, I dont see anything after Coomassie stain?

I can see it with Silver stain after coomassie but the stain is very poor. However, In total serum I can see proteins with Comassie Stain. the LMW proteins were obtained by ultrafiltration on 20...

19 October 2015 9,230 4 View