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Questions related to Protein Expression
I want to clone a target gene into a lentiviral vector to produce a mouse cell line that overexpresses the target gene. I want to know the difference between inserting the full-length cDNA and...
07 November 2019 4,499 2 View
Hey everyone, I made bacterial cell lysates and blotted them to check virulence factors. The two virulence factors I investigated worked quite well and specific bands were visable. However, now I...
16 October 2019 6,703 3 View
Hi, I am looking at a change in protein expression after treatment over time and comparing samples to an untreated control. However, certain proteins i.e. stress response proteins, does not...
10 October 2019 3,740 4 View
I'm looking for a nuclear extraction kit, and upon searching, every vendor offers one with each kit seemingly the same. What kit do you use, and why do you like it over the multitude of...
10 October 2019 2,933 1 View
I am studying the expression and toxicity of a GFP construct using pBAD24 on E.coli BW25113. First I used LB as a medium, but due to heterogeneous expression between cells I decided to use...
10 October 2019 1,616 3 View
I have transfected the HEK 293T cells with plasmid having fluorescent protein expression. After sorting the positive cells (cells having fluorescent protein expression), i want to isolate the...
19 September 2019 1,912 3 View
09 September 2019 5,022 3 View
Hi all, I want to download a gene sequnce from Genome Browser, but I am not so sure about the direction of the sequence I get. Does it always show the coding strand sequence of one gene in the 5'...
06 September 2019 6,242 2 View
I'm trying to analyze how the ratio of AMPKp: AMPK changes among my treatment groups. What is the best way to do this? Currently, all I've done is divide AMPKp by AMPK, and then calculate...
31 July 2019 5,234 3 View
I want to use E. faecalis JH22 as competent cells to express my protein using anhydrotetracycline (ATc) to induce protein expression i need some standard protocol if someone has use it before.
27 July 2019 2,092 2 View
07 July 2019 1,974 4 View
07 July 2019 9,566 2 View
I am looking at different methods to measure the expression of proteins in vivo in plant cells. I know that GFP-tagging is the most historically common, but I'm just brainstorming (so don't worry...
07 July 2019 6,939 6 View
Hi everyone. I have to assess protein expression in my adult satellite muscle cells by WB. I'd prefer not to use the conventional beta-actin or alpha-tubulin, nor GAPDH. I saw that 18S mRNA was...
25 June 2019 3,301 2 View
I am going to boldly attempt the purification of a viral membrane protein and want to try several expression systems to see what works best. One I would like to try is pBAD because (in my hands...
19 June 2019 3,970 1 View
I am looking to measure phospho-Erk and total-erk protein expression with western blotting, and am wondering if it is necessary to use a loading control. I normally measure phospho first, then...
12 June 2019 8,318 2 View
06 June 2019 7,885 1 View
Hi, I have produced a 37kda viral protein using Baculovirus expression system (FlashBac Ultra)/insect cells. It is a glycosylated protein and contains a transmembrane region at N-terminal....
06 June 2019 6,641 8 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...
29 May 2019 3,045 2 View
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!
13 May 2019 441 0 View
If there was a very strong RBS site placed between the promoter and the AUG of the fusion partner at the N terminal, while there would also be the Shine-Dalgarno sequence placed in between the...
05 May 2019 2,478 4 View
05 May 2019 9,647 3 View
05 May 2019 7,001 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 9,500 3 View