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

More Prabhadevi Venkataramani's questions See All

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...

11 December 2015 2,805 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...

07 August 2014 5,925 7 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...

06 July 2014 3,591 14 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...

06 July 2014 3,947 6 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...

06 July 2014 9,903 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...

06 July 2014 7,793 9 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Nawroz Abdulrazzak

Use Histidine phosphatase which catalyze core centred on a histidine which becomes phosphorylated during the course of the reaction

Manoj Balakrishna Menon

According to the company Lambda phosphatase from NEB is effective for dephosphorylating phospho-Histidine. I have not heard of anything commercial, if u need ''only'' histidines to be dephosphorylated