Barney .

10 Questions 14 Answers 0 Followers

Questions related from Barney .

Why is it that we can't quantify cDNA?

I know this is probably going to sound like a stupid question but why is it that you cannot quantify cDNA, as I've been told? Or is there? Just looking for some insight. Cheers, Mat

22 August 2017 6,525 1 View

Why is it that we can't quantify cDNA?

I know this is probably going to sound like a stupid question but why is it that you cannot quantify cDNA, as I've been told? Or is there? Just looking for some insight. Cheers, Mat

08 August 2017 4,310 1 View

What's the best mounting medium w/ DAPI?

Hi, I am doing in situ zymography to measure MMP activity in heart tissue sections. I am trying to decide between different mounting media w/ DAPI. 1. VectaShield Antifade w/ DAPI (H-1200) 2....

08 August 2016 7,075 7 View

Can FKBP proteins create a complex with a mammalian expressed protein?

I am currently trying to purify a protein called MMP-2. It has an approx. molecular weight of 72 kDa. For the past several weeks, I have been purifying this protein expressed from BL21-RIL E.coli...

12 December 2015 5,215 6 View

What happens if the Ni-NTA resin on my column runs dry?

Hi, I am doing an affinity chromatography protein purification for a protein of interest. I made the error of leaving my pump on too long and my buffer solution ran dry and made my column run dry...

10 October 2015 3,080 9 View

What happens if the Ni-NTA resin on my column runs dry?

Hi, I am doing an affinity chromatography protein purification for a protein of interest. I made the error of leaving my pump on too long and my buffer solution ran dry and made my column run dry...

05 October 2015 7,593 7 View

How can I tell if my protein is still bound to my resin?

I am doing IMAC protein purification with a his-tagged protein but I have been having issues purifying my protein. First, the first few times I did it, I noticed that much of the protein was found...

08 August 2015 3,860 4 View

What's the best way to maximize protein binding to my column/resin in order to purify a protein with an Ni-NTA column?

I have done 2 purifications using a Ni-NTA column so far and each time we've noticed (based on my SDS-PAGE results) that lots of protein is collected in my flow-through (not binded to the...

27 July 2015 844 14 View

What's the best way to maximize protein binding to my column/resin in order to purify a protein with an Ni-NTA column?

I have done 2 purifications using a Ni-NTA column so far and each time we've noticed (based on my SDS-PAGE results) that lots of protein is collected in my flow-through (not binded to the...

07 July 2015 821 14 View

Will my pre-equilibrated Ni-NTA column be compromised if I run 20% ethanol through it? Will have to re-equilibrate my column with the appr. buffer?

I am doing a Ni-NTA chromatography purification of my protein. I noticed that air bubbles get caught within the column when I've poured my resin into it and I read somewhere that I can use 20%...

07 July 2015 7,719 9 View