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 need these bacterial strains for my research
28 December 2019 6,822 0 View
Can any one suggest me concentration of beta-mercaptoethanol (2-ME) or DTT to prevent cysteine dimerisation in protein purification at pH 8.0 . and stability of them in Tris Hcl buffer with...
17 December 2019 3,120 4 View
12 December 2019 6,133 4 View
I’m still a little new to this technique [of purifying his-tagged proteins]. I know there are protocols for recharging the columns but does the resin need to be removed before purifying the next...
12 December 2019 6,088 7 View
Does anyone have experience trying to work around background fluorescence in cyanobacteria? I'm currently trying to image a GFP fusion protein, but I didn't realize there was significant...
05 December 2019 5,933 3 View
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...
15 November 2019 6,192 14 View
Hi All, I am currently working on a protein-miRNA binding assay. I am using FP as my assay system to see if the binding happens. Previous publications show the protein binds to the miRNA and my...
14 November 2019 8,768 2 View
I am working on a membrane protein of size 70 kDa. It has a 6X-His Tag on C terminal. I am using 20mM Tris, 200 mM KCl, 10% glycerol, 5 mM beta-mercaptoethanol and 0.03% DDM for the protein...
11 November 2019 1,318 3 View
Dear All, I am using Anti-Flag magnetic beads for immunoprecipitation experiments. I want to perform elution under the non-denaturing conditions for further use of proteins as well as to reuse...
11 November 2019 4,886 7 View
11 November 2019 9,540 2 View
11 November 2019 7,084 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 5,302 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 2,213 3 View
Several papers have cited chicken pancreas and dessicated hog kidney as the source for folate conjugase enzyme. 2 ml of this added at a concentration of 5mg/ml. Kariluoto & Piironen (2009)...
25 October 2019 9,182 0 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 6,683 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 2,211 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 5,447 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 7,751 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 8,598 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 5,656 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 5,639 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 949 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 6,498 3 View
10 October 2019 4,128 4 View