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, Recently, I purified 8 proteins and stored at -20 with the elution buffer containing 20 mM Tris, 250 mM NaCl and 200 mM Imidazole. I autoclaved the buffer, so I do not know whether the...
26 September 2020 10,238 6 View
I asked for protein purification, identification and isolation of unknown proteins and test versus GS-9L other than that we already got in our patent WO/2014/040605 - form native sample of honey?!
24 September 2020 2,206 3 View
Hi all, I have a purified protein (57kDa) which tends to precipitate at 4 deg C after a week (conc. 9mg/ml). Can you please share the protocol to flash freeze protein in Liquid nitrogen and...
17 September 2020 1,728 5 View
i want used COLLAGENASE TYPE IV for isolate SYNOVIAL FIBROBLAST in cartilage. but the tissue is too small to isolation and i don't detect part of cartilage so lsolated from rib and cartilage...
16 September 2020 2,129 0 View
As Chenopodium quinoa is tolerant to wide range of salinity stress. i want to carry out RT-qPCR for C.quinoa under salinity stress. so which gene should i use as marker/control in quinoa expressed...
05 September 2020 9,665 3 View
Hello all, I frequently use the Akta purifier for protein purification and recently have been getting error messages in unicorn saying "error 2012 no response frac, 900". The frac 920 is designed...
03 September 2020 3,032 1 View
I need to concentrate DNA using filtration based purification methods. Between vivaspin and microsep which one is better for more DNA concentration output.
15 August 2020 8,431 1 View
Hi :D I'm doing purify a small protein from E. coli. From E. coli extract supernatant, I purified my protein using IMAC purification with LC. I got a single peak at chromatogram and got the...
24 July 2020 2,710 5 View
I'm purifying TB antibody and the columns we are using only seem to have a DBC of about 0.15 mg/ml. This seems very low to me, but I am a protein purification novice. Is this normal?
15 July 2020 1,645 4 View
I am trying to purify a plasma membrane protein from ecoli. I think the standard procedure for membrane protein extraction is like this: Lyse Low speed spin to clarify (eg, 5k xg for 20...
03 July 2020 6,184 3 View
I need to find the number of copies of a specific membrane protein per E. Coli cell . I've created different mutations of this proteins and I need to figure out how would these mutations affect...
01 July 2020 9,179 0 View
I am working with an unusual bacteria that is very hard to lyse. I want to extract protein and RNA from this bacteria. I need to determine the optimal lysis conditions for this bacteria, but I am...
28 June 2020 5,196 0 View
Unfortunately, our stock of Lipofectamine 2000 got contaminated by bacteria: how can I sterilize it without losing their DNA transfection properties? Do you have a Dos and Don't's list? Thank...
18 June 2020 4,165 13 View
Hello! I have some expired Ni-NTA resin (QIAGEN), and I need to purify my His-tagged protein. Does the expiration date affect the binding capacity of Ni-NTA? Thank you in advance
12 June 2020 6,086 5 View
I have a question about protein transfection. I used a reagent to transfect a protein. Can I use the same reagent for a different protein that I want to transfect in the same cell line?
01 June 2020 2,794 3 View
From my experiment, I determined MDA, AST, ALT, SOD, T. Cholesterol and HDL Cholesterol in various organs Plasma, Liver, Kidney, and Heart. To get their relationships in each of the organ, I drew...
23 May 2020 7,179 0 View
Hi, I purified a protein (about 28KD) via the following steps: 1- Firstly, I used buffer A native(Qiagen)-NaH2PO4,NACL,Immidazol,ph8 2-then after centrifuging and separating my pellet , I added...
14 May 2020 1,890 6 View
Hi, I'd like to have an advice on which buffer and/or procedure I should use to maintain the phosphorylation status of a protein to be analysed through mass-spectrometry. I have to lysate breast...
11 May 2020 2,167 4 View
The proteins need to be enzymatically active, planning to use Taq. Polymerase for PCR reactions. I am looking to create a protein extract that can be used for diagnostic PCR. So the protein has to...
11 May 2020 7,388 12 View
Dear all, I'm looking for Cpf1 purification protocol and I found a protocol in which the authors added GABA to the buffers ( from equilibration till the eluition). I'd like to know if I can...
04 May 2020 7,841 6 View
In measuring the plasma HDL-c levels by a phosphotungstate-magnesium method, and the following enzymatic measurement of cholesterol in the supernatant, if I run out of the kit's standard...
01 May 2020 388 0 View
I have got multiple categorical variables which are all binary (yes/no). I would like to visualise 3-4 variables against one predictor variable in the same graph.
29 April 2020 1,690 0 View
We are expressing B-amyloid peptids and after purification we are having some problems with the aggregation of the samples. I'm wondering if there are optimal conditions of storage that dont...
08 March 2020 9,604 3 View
I'm extracting protein from highly hazardous bacterial samples. For samples like this, people commonly boil the bacteria prior to full lysis. Because the bacteria is boiled, do I still have to...
03 March 2020 6,155 3 View