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 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...
06 October 2019 5,008 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...
03 October 2019 9,602 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...
01 October 2019 3,628 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 466 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 5,472 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 6,914 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 9,792 6 View
09 September 2019 553 5 View
09 September 2019 8,336 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 6,583 4 View
09 September 2019 8,413 7 View
Hi everyone, The DE-52 column is equilibrated with [pH 8.0 20 mM Tris-HCl, 1 mM EDTA] buffer, and the pH is around 8.5 at 4 deg (checked with pH meter). The Mb (pI ~ 7) is dialysed overnight...
09 September 2019 7,652 10 View
I'm attempting to concentrate whole cell lysates from primary murine cells, in order to load enough protein onto a gel to detect my protein of interest. I'm pretty new to protein purification and...
09 September 2019 7,593 3 View
hi, i'm doing a protein purification and i'm wondering what are the factors make a single SEC peak give many bands in a BN-PAGE.
09 September 2019 7,615 8 View
Hello everyone, I try to purify a chitin binding domain in a native conformation in mono disperse suspension. After purification by use of a polyhistidine tag I dialyze the protein suspension in...
14 August 2019 2,928 6 View
Hello all, I am currently reading up on clear coli BL21(DE3) cells as a possible alternative to BL21(DE3) cells for recombinant protein production such that my end products are not contaminated by...
08 August 2019 2,949 3 View
08 August 2019 2,803 7 View
I am trying to concentrate enveloped RNA viruses from infected cell culture supernatant. The clarified media (MEM containing 0.375% BSA fraction V.) will be mixed with PEG8000( to a final...
03 August 2019 407 3 View
I'm trying to express recombinant protein from mammalian cell 293F by its secretion in to the supernatant and then using the Ni-beads to enrich the His-tag protein. However, when I use 300mM...
11 July 2019 4,662 5 View
I have seen lots of papers in literature using prep cell for purifying nucleosomes (from DNA and hexasome species), but I couldn't seem to find another protocol apart from that discussed by Luger...
07 July 2019 6,816 4 View
07 July 2019 2,120 6 View
Is there a more stringent elution buffer than 2% SDS with BME? I have tried this both at RT and 50 degrees with very little antibody eluted on SDS-Page. Based on ELISA, the antibodies are...
07 July 2019 3,009 4 View
I am attempting to purify a protein with his-tag. Current lysis method includes his-lysis buffer (pH8) with 1% triton and 2minute sonication (pulse 10seconds, rest 40 seconds) after incubation...
07 July 2019 2,986 10 View
I have a set of buffers (Tris, imidazole, phosphate) for NI-NTA protein purification. I prepared them about three weeks ago and they contain DTT and PMSF. Can I add fresh DTT and PMSF to the...
07 July 2019 3,467 4 View