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
Hi all, I have this his-tagged protein which I wanna pull down and want to perform a Chip-seq. While Ni-NTA is used for a lot his-tagged protein purification, I didn't find any citations using...
10 October 2018 8,151 4 View
I have get insolube protein at 1mMIPTG after 4 hours induction time. How can I find solube protein for purification by his tag nikal colum.
10 October 2018 8,437 6 View
I am trying to purify a membrane protein and a transcription factor. We are planning to use Step tag for purification. Can someone shed light on this method? Is it better than traditional GST and...
10 October 2018 5,617 0 View
Dear friends, We have a bunch of small molecules (around 20) and purified protein as potential target. I am wondering what is the fastest and cheapest experiments to find if there is any...
10 October 2018 3,711 28 View
Hi guys, I am wondering if anyone has using TB instead of LB for the pre culture of isotope labeled protein (C13 and N15) and what the yield is like compare to LB transfer. More precisely, my lab...
10 October 2018 7,226 4 View
03 October 2018 4,597 27 View
I am doing experiment to produce alpha glucosidase from thermophilic bacteria. Though I can determine enzyme activity by enzyme assay but my supervisor wants me to have a positive control, in this...
09 September 2018 3,452 11 View
I extracted rice leaf (fresh leaf) total protein using following protein extraction buffer on ice bath or at Room temperature (without liquid-N2). After that, I did Western blot (WB) to check my...
09 September 2018 4,669 8 View
I am trying to purify a N-terminal his tagged secreted protein. The protein of interest is a self assembled nanocage and is engineered to express his-tag on its surface which enables downstream...
09 September 2018 8,136 4 View
I've worked in both labs that used sonicaiton vs french press to lyse cells. The lab im currently in doesn't have a french press, and some protocols noted the use of french press. I was wondering...
08 August 2018 5,284 1 View
I am going to extract protein from plant leaf tissue using 2% PVPP (Polyvinylpolypyrrolidone, 110 μm particle size). But this PVPP is insoluble in water, means in my protein extraction buffer....
08 August 2018 6,411 9 View
I am constructing one plasmid vector for mammalian expression. I am going to use it for Purification of Protein for structural analysis. Which Promotor is the best one and can give me high yield?
08 August 2018 734 1 View
Hi everybody, I'm currently expressing proteins in and extracting from bacteria. The procedure is that I pellet the bacteria from bioreactor, resuspend in buffer (50 mM Tris/HCl, pH 8.0; 1M NaCl;...
08 August 2018 2,563 9 View
My insect cell pellet (GPCR expression by using baculouvirus) has been freezed and thawed twice. Is it still good for GPCR protein purification?
07 July 2018 8,702 3 View
Our lab uses a lot of Ni-NTA beads, and our small gravity columns mean it can take several hours to clean/regenerate them between different recombinant proteins. I am wondering if it is...
07 July 2018 5,788 2 View
Hi I want to purified a protein for BL21 by sonication. i need the new protocol about that thank you
07 July 2018 7,491 1 View
Dear colleagues, do you have any suggestion how to get rid of high molecular weight proteins from blood serum? I do zymography and bands of these proteins (>135 kDa) stay coloured dark blue. It...
06 June 2018 7,780 5 View
I want to separate protein from a formulation containing glycerin, phenol, metacresol, zinc, disodium hydrogen phosphate and NaCl. Can a desalting column like Zeba (Thermo fisher) or Amicon...
06 June 2018 4,393 6 View
Dear All, Sonication is a vital step in protein purification and over sonication can definitely damage the secondary structure of the protein(correct me if I am wrong). What is the best Hz value...
06 June 2018 9,168 6 View
Good time of the day, I need to perform co-IP (and X-ChIP with the same targets and antibodies) For co-IP i used Pierce IP/co-IP kit (#88805). E.g. that one that uses antibody crosslinked to...
06 June 2018 7,953 5 View
are all proteins are precipitated by cold acetone? how can acetone be useful in protein purification?
05 May 2018 1,355 0 View
Hello all, I am a Master student and in my M.Tech project, I am working on Chikungunya virus. I am trying to purify it's envelope protein. i have used affinity chromatography (His tag, Ni-NTA,...
05 May 2018 6,286 6 View
No explaination
05 May 2018 8,139 3 View
I have a 28kD protein and am trying to get rid of a 10-12kD contaminant using sephadex-25 medium. I was wondering the column size, volume and buffers that would be best for this since I am very...
05 May 2018 4,693 8 View