Prabhadevi Venkataramani

7 Questions 10 Answers 0 Followers

Questions related from Prabhadevi Venkataramani

What transfer conditions should I use to blot a membrane protein of size 82kDa?

I need to transfer a membrane protein (size 82.9kDa) onto a PVDF membrane. Can anyone help me with the transfer conditions I need to use for this? Is 70V for 4 hrs enough or should I transfer even...

12 December 2015 2,883 3 View

What kind of post translational modifications can cause a huge change in the molecular weight of a protein with the exception of glycosylation?

I have run a 2D gel to identify certain protein isoforms in rat urine. I did a western with a antibody that binds to all isoforms of the protein. The expected size is around 17kD-20kD., however...

08 August 2014 5,966 7 View

How do I remove phosphate group from a protein that has been phosphorylated on its Histidine residue?

I wanted to know if there is a phosphatase that is specifically used for removing phosphate groups from histidine residue of my phosphorylated protein. Mostly, I have only heard of Ser/Thr...

08 August 2014 9,780 2 View

What are the membrane transfer conditions I should use during western blot if i have two proteins of varying sizes?

If i need to do western blot of two proteins of varying sizes separated in the same gel (one 55kDa and one 13kDa), what should be the membrane transfer conditions that I need to use? Is 70V for 2...

07 July 2014 3,984 6 View

How can I solve my protein instability so it can be used in NMR and ITC studies?

I have a highly charged & low molecular weight bacterial protein that is highly unstable. I have tried purifying it with different tags including His tag, MBP tag, Thioredoxin tag, SUMO tag...

07 July 2014 3,631 14 View

Does the use of Phos-Tag give a high background?

I will be incorporating Phos Tag in my protein gels to study histidine phosphorelay between two bacterial proteins. I wanted to know if there is a possibility of getting a high background upon...

07 July 2014 9,947 2 View

How to check protein stability?

I have purified my protein and I want to check its stability. If I keep a tube with my purified protein at 4C for a day, can I check its stability at the end by spinning it down and measuring OD...

07 July 2014 7,856 9 View