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Questions related to Protein Expression
Actually , i want to express Thermomyces lanuginosus lipase (TLL) in E coli system. so please help me and if somebody have protocol/procedure plz share with me. Thanks in advance!
05 May 2019 9,759 3 View
Hi All, We are interested in using the Branson SFX150 sonicator we have in our lab to lyse DE3 E.coli in order to purify a couple of proteins. I have worked with larger sonicators in the past to...
05 May 2019 7,078 2 View
Hi, guys, I'd like to screen a receptor for one protein, so that I need to make every membrane protein express on 293 cells, but I have no ideas for getting these expression vectors. I had...
05 May 2019 9,546 3 View
Hi everyone, I have to express different mammalian proteins tagged with different fluorophores (such as GFP, YFP and others) under bacterial expression system and to use these purified proteins...
25 April 2019 1,653 3 View
I am currently working on several proteins with at least 4 disulfide bonds. I am isolating inclusion bodies and refolding the proteins. I would like to try expression of the same using CyDisCo...
22 April 2019 1,570 3 View
Hi everyone! Previously, I transformed a BL21(DE3) strain with a pET-15b vector carrying my gene of interest. The strain grew and I could express my protein. Now I need to do that again, but...
22 April 2019 6,659 4 View
I'm working with E. coli for protein expression and I'd like to speed up the process if possible. The standard protocol is to add a 1:100 ratio for starter culture to the larger volume (ie. 10mL...
04 April 2019 7,028 4 View
Hi, I am working on Pichia pastoris fermentation. I had runned on 7.5 litter scale, small scale. I have yet tried at 25 litter scale, big scale. The result shows I got failure in scaleup. You can...
04 April 2019 3,583 10 View
04 April 2019 1,653 4 View
04 April 2019 3,060 5 View
04 April 2019 630 4 View
I tried to downregulate one protein expression in RAW264.7 Macrophage cell line using recommended RNAiMAX Lipofectamine, but not geting any good result. I tried different siRNA sequence already?
04 April 2019 7,595 2 View
sometimes the protein can be expressed even without adding iptg, so is it necessarily to add iptg for protein expression?
04 April 2019 7,745 2 View
My protein is transmembrane ion channel, and I want it to be functional in membrane. Here I think that the plasmid should contain a signal peptide that express in frame with my protein and guide...
04 April 2019 7,056 1 View
I’m looking for the best available expression system that can produce fish protein. The downstream application of the recombinant protein is functional analysis , enzyme activity assay,...
22 March 2019 6,743 2 View
I am studying about recombinant protein to produce vaccine, and I use pET32a as vector for protein expression. I wanna ask are there any effect of the thioredoxine tag (Trx-Tag) in the immune...
03 March 2019 1,466 3 View
03 March 2019 6,997 2 View
I have sequence for Factor Xa site in my expression vector, which will get translated to Ile-Glu-Gly-Arg after protein expression. What is the purpose of this site?
03 March 2019 5,955 1 View
Hello, I have cloned a gene coding for a non-secreted proteolytic protein into E. coli BL21-AI and I am looking for an easy way to lyse these cells in order to extract/purify the protein. I know...
03 March 2019 6,751 4 View
While studying a particular pathway under an induced metabolic state, I have noticed that the protein expression levels often increase in 6 hrs, decrease in 12 hrs and then subsides to normal...
03 March 2019 3,557 5 View
The pichia pastoris was GS115, and the vector was pPIC9K. The previous steps including electrotransformation, His- plate selection and G418 screen were normal. However, when I ready to express...
03 March 2019 4,388 7 View
Hi, I found these little circle caps/membranes(?) in my lab, they are stored in an odourless liquid that is not water (I tried mixing some in and could see the fluids mixing). I work in a...
20 February 2019 7,547 7 View
Hi Everyone Our group wants to compare protein expression levels in human serum samples. So we want to do some proteomics to get the list of differentially expressed proteins in those samples....
02 February 2019 2,911 4 View
02 February 2019 6,967 8 View