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Questions related to Protein Expression
How to measure the diameter of plaque
22 February 2022 3,414 4 View
Dear community, I want to use the Friedman-test in combination with the Dunn-test on my data in GraphPad Prism. However, I have some missing values, so the software is not able to perform the...
24 January 2022 8,813 1 View
Generally, people can use either deletion or overexpression strategy to do characterization of any novel protein candidate. Can anyone describe or suggest any reference paper discussing the pros...
22 January 2022 9,152 4 View
I have purified a recombinant protein and I am not getting any protein activity in activity assay. Protein contains histidine tag. Is there possibility that his tag is interfering with protein...
14 December 2021 8,152 4 View
Dear everyone, I am trying to express the extracellular domain of a membrane protein in HEK 293F cells. The construct contains the signal peptide of the protein itself. However, the protein does...
26 November 2021 3,874 7 View
I'm designing a construct with iCre. The sequence from the publication describing this modified Cre sequence for the first time (Shimshek et. al 2002, genesis, vol. 25;32;, no. 4;1;, pp. 19-26)...
23 July 2021 2,043 1 View
Hi, I am working with pEGFP-N3 SV40 origin containing plasmid to assess transfection efficiency in different cell types. I am struggling to compare the efficiency of transfection between two cell...
04 July 2021 8,477 0 View
I use vcftools to calculate FST between different populations, based on resequencing data. But the values of FST are between 0.5 and 0.9. My research object is an endangered plant. I want to know...
21 June 2021 7,396 1 View
I am seeking to use in vivo injection of AAV in adult mice to express TEV protease in to the heart muscle cells of our mouse line. We do not have experience with this, but based on the literature,...
26 April 2021 9,297 1 View
Hi there, I am trying to purify a recombinant His-tag protein from insect cell (baculovirus expression system) I used buffer containing %10 glycerol, 50mM SODIUM PHOSPHATE, 300mM NaCl , 20mM...
19 February 2021 6,941 6 View
i have issues in expressing a chicken protein in BL21 cells with his tag. I do not see induction even at 18 degrees. Any suggestions
16 February 2021 9,125 3 View
I am using KRX cell. I have grown my overnight cultures at 37 degrees. When I inoculate 1L TB media, my cells do not grow. I have also tried using 25 degree incubation of overnight cultures and...
11 February 2021 4,029 3 View
Are E.coli strains M15 and pET28a vector systems compatible? Can I over-express my protein which is of plant origin. Previously I was using BL21 DE3 strains. Was confused to use M15 strains for...
11 February 2021 2,808 3 View
I did a Co-IP MS with two samples and got the LFQ data from MaxQuant, and I wanted to normalize it by an internal reference protein. After I divide the LFQ of the other protein by the LFQ of the...
19 January 2021 5,628 4 View
Dear all, When I run my yeast total protein lysate extracted after 10 minute-treatment with my compound, I observe a global up regulation in proteins, even my loading control protein. I suspect...
04 January 2021 1,527 8 View
Hello, I've never worked with fiber protein before and was hoping I could purchase it online vs creating it in the lab. I cannot seem to find a protocol for ad5 fiber protein expression, so...
23 December 2020 2,023 2 View
I ran a Bradford assay and also used direct UV absorption to find the protein concentration of my UVRBprotein sample. The latter gave a much higher result- almost double that of the Bradford...
20 December 2020 1,056 2 View
We recently trying to express scfv-his tagged through Cytoplasmic expressions using SHUFFLE strain. We are getting a high protein expression related to 50 kDa, instead of ~27 (our target). This...
19 December 2020 4,795 1 View
The memory error "stack overflow" came out when I used Autodock vina to choose flexible residues. I only use a ThinkPad PC with i7 8th Gen processor. Could anyone help me with this question....
11 December 2020 4,855 2 View
Hi I've been trying to produce some recombinant human Interleukin 6. I sub-cloned the gene for human IL6 into a Promega T7 Flexi vector (native gene no tag yet) and confirmed sequencing and...
10 December 2020 2,532 4 View
Hi I need to purify a recombinant protein (IL6 21kDa). It's been produced in E.coli BL21 with a T7 expression vector, but it does not have a tag as yet. I was thinking of adding a his-tag and...
10 December 2020 2,384 8 View
I don't believe there's much research in this area in humans. There are mice and primate studies
06 December 2020 4,666 2 View
Hello all, I am currently trying to modify my plasmids so that it has a sequence coding the pelB or ompA tag in front of the proteins. The problem is that in order to paste such a sequence into...
26 November 2020 8,527 3 View
I am expressing a construct in E. coli under different conditions to check soluble expression. I have a large array of conditions, which cannot be done in flasks. Thus, I will be growing cultures...
02 November 2020 2,837 6 View