Contact experts in Protein Expression to get answers
1,932 views 1,475 posts
Questions related to Protein Expression
only WB is sufficient along with mRNA expression to present the data without IHC??
15 April 2020 5,728 4 View
Hi, I am currently using BL21(DE3)PLySs for protein expression. In our lab, we are using selective antibiotics for both inoculation(small culture, a.k.a over night seed culture) and scale up(Large...
07 April 2020 2,828 9 View
Hi fellow RGers, in my experiments, I'm trying to express a peptide and I'm curious about the general stability of the expressed peptides. Is there a rule of thumb concerning peptide stability in...
26 March 2020 9,919 3 View
I could not really find a protocol specifically stating how to induce protein expression under AOX1 promoter in Pichia pastoris. Do we straight away streak them out on BMMY agar? Or streak them...
16 March 2020 9,873 3 View
Hi everyone. I am trying to express and purify a recombinant antimicrobial peptide using as a host cell E. coli BL21 (AI) (employing L-arabinose and IPTG). Because this peptide can be toxic to...
20 February 2020 5,175 4 View
I'm trying to inhibit a signalling pathway in a breast cancer cell line (MDA-MB-231) through overexpressing its physiologic antagonist via adenoviral infection. The problem I'm facing is the...
14 February 2020 3,551 0 View
Does anyone know of a more affordable alternative to F17 media for 293Expis? We currently supplement with L-glutamine and Pluronic F-68.
11 February 2020 796 1 View
In line with previous data suggesting low efficacy of the CMV promoter in some cell lines (Qin et al. PlosOne 2010), I experience low transcriptional activity in a neuroblastoma cell line after...
11 February 2020 2,380 3 View
02 February 2020 5,984 4 View
Hello. I want to crystallize kinase domain of a protein in active form. The active form of the domain has specifically phosphorylated tyrosine residues. However, I am not sure if I can do so in a...
02 February 2020 7,604 5 View
It seems to me that cell-free protein synthesis is not very used when it comes to protein expression, though its usage should avoid many problems, like the presence of inclusion bodies. I was...
02 February 2020 7,700 3 View
Hi everyone, I am a newbie in molecular biology. Recently, I encounter an issue that the non-uniform fluorescence protein expression level in lipofection transfected HeLa cell. The variation would...
02 January 2020 7,330 4 View
I'm implementing a small microbiology lab and requiere an homogenizer for the meat samples we produce. A small hand blender will sufice, since the amount of yearly samples will be small, hence a...
02 January 2020 1,564 1 View
Hello Experts, I am planning to do the protein expression in bacteria. but my lab has only BL21 (CDE3). I have been searched a lot, but I couldn't find any information. Can you tell me the...
01 January 2020 1,423 3 View
I have a mixture of plasmids (with Origin of replication that can work in Pichia pastoris) with different genes for protein expression. Is it possible that after chemical heat shock...
22 December 2019 5,788 3 View
Our truncated protein expressions (labeled by the HA tag) in lane 4 and 5 were supposed to be matched with the mRNA expressions in lane of 293FT-4 and 293FT-5. However, the different truncated...
20 December 2019 3,439 7 View
12 December 2019 5,445 7 View
Dear all, I checked my protein expression in the sphere cells through western blot. I could not see any expression. I added whole cell lysate and sphere cells. I could not see any expression of my...
12 December 2019 8,564 3 View
Hey, I did WB and IF in different time but with the same cell line, however, the results from WB and IF seem incompatible. WB and IF both show the protein expression level right? How could they be...
12 December 2019 6,537 4 View
I was successful with the transformation( pPIC9K vector in Pichia pastoris GS115 strain )and got good colonies in RDB plates after 48 hours. later in MD and MM plates also. Can anyone please tell...
12 December 2019 8,966 8 View
10 December 2019 6,556 8 View
Hello all, We have a discussion in our lab and would love your input: Is it possible for cultured bacteria that are in, or approaching, their stationary phase (in our case OD 5 - 6) to go back...
10 December 2019 1,315 8 View
This is the upper part of a western blot membrane, and as you can see there are some proteins that did not enter the separating gel. Does this affect the level of protein expression? I mean is...
16 November 2019 1,487 4 View
I need to grow my bacteria which is Streptomyces sp. from spore suspension method. I have grown them onto the nutrient agar by spread plate method and now I incubate them for 5 days. And for the...
15 November 2019 1,194 3 View