A forum to address questions regarding methods of protein purification. | Contact experts in Protein Purification to get answers
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Questions related to Protein Purification
I have a major problem that I have been facing for last few months. I have a protein which is His-tagged at the C-terminus. Its a fusion of 5 monomers and its weight is around 42 KDa. The...
11 November 2019 7,073 14 View
I am currently having difficulty optimizing a binding buffer for Ni-NTA protein purification under denaturing conditions. My starting buffer is currently composed of 100 mM NaH2PO4, 1 M NaCl, 10...
07 November 2019 5,288 3 View
What are the current methods to quantify, measure, and detect the content of Immunoglobulin G proteins in human blood? How much time does it take for such tests to complete? How much does it take...
25 October 2019 2,198 3 View
Hello everyone, I am currently working with targeting proteins to yeast mitochondria. Our strategy is to use a mitochondrial targeting signal to transport our protein of interest to mitochondrial...
17 October 2019 6,664 5 View
Hello, Any one have a protocol for disrupt cells and obtain cytoplasmic proteins ready to load in HisTrap HP columns? I'm using transient transfection of HEK2913T for recombinant protein...
10 October 2019 2,199 3 View
I want to resuspend a lyophilized protein in buffers that mimick these cellular compartments. Do you know any references about that? I guess I should use a crowding agent but it should be...
10 October 2019 5,429 2 View
In the lab I am working in, we are growing cells 3h, 37°, inducing them with IPTG when OD is nearly 0.6. After 3h, they are harvested for purification. Usually they take 80-90 minutes to reach...
10 October 2019 7,741 4 View
If you need to filter 500 ul of a protein that´s in a buffer with 1% DMSO, how would you filter this before putting in an analytical equipment, like for SPR? Would you use regenerated cellulose...
10 October 2019 8,591 6 View
I am looking into doing a peptide conjugation and the protocol im looking at says to use dialysis tubing to separate the reacted peptide from unreacted. I do not have much of the peptide so i...
10 October 2019 5,634 3 View
I want to buy chromatography columns for the affinity-based separation of proteins. The size (diameter) of the column needs to be very small (less than 100um) in order to match the design. Are...
10 October 2019 5,624 5 View
Hi all, I'm trying to purify an over expressed MBP tagged protein. I had a hard time expressing it in E. coli and moved into Sf9 insect cells. I've managed to overexpress the protein, but when I...
10 October 2019 937 14 View
Base on my understanding, PMSF is an inhibiror for serine protease. I notices that some proteins (eg. ana Fld) requires PMSF in its purification process, usually during the sonication step. Other...
10 October 2019 6,485 3 View
10 October 2019 4,110 4 View
06 October 2019 4,974 4 View
03 October 2019 9,584 5 View
01 October 2019 3,606 3 View
I am cloning a gene whose protein product is endonuclease what enzyme assays should I do after successful purification of the protein
30 September 2019 433 7 View
I am planning on purifying a small 12 kDA protein with a 6x-His tag using Ni-NTA resin. Many protocols recommand the addition of protease inhibitors like PMSF for optimal lysis and purification of...
20 September 2019 5,434 4 View
Do we need to place the insert in a specific site in his tagg vector for further protein purification or can we just use any restriction site present in the vector?
19 September 2019 6,892 3 View
We have a cellulase crude protein mixture with oligosaccharide contamination. I have tried various protein purification methods like Ammonium sulfate precipitation, Column based, TCA method,...
17 September 2019 9,768 6 View
09 September 2019 520 5 View
09 September 2019 8,313 3 View
I need IPTG to induce big volumes of E.coli cultures to do protein purification. Many suppliers advertise their IPTG as being "dioxane-free". What difference does it make ?
09 September 2019 6,552 4 View
09 September 2019 8,394 7 View