Currently wanting to look at chromatin-bound vs soluble protein concentrations.
My PI was at a conference this spring where they used the intensity of the DAPI stain of fixed cells to be able to tell the cell cycle phase of individual cells in an asynchronous population. I'm...
06 July 2016 8,571 0 View
Hi all, I am wondering as to the conditions of a lysis buffer that would lyse both the cell membrane and the mitochondrial membrane. The purpose of this lysis is to collect genomic and...
09 October 2014 514 3 View
I'm looking for a lysis buffer recipe and protocol for lysing an E. coli strain. The background is that I will be using IPTG to induce the expression of a plasmid with a specific cDNA. I...
10 November 2013 6,245 6 View
I am currently staining cells to quickly look at the effect of a drug for both 24 hour and continuous treatment. At the end of the assay, the cells are to be stained with Giemsa and will be judged...
31 December 2012 3,059 8 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...
06 August 2024 5,801 4 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View
I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...
28 July 2024 366 3 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
It has been long known that EDC carbodiimide and NHS easter's half life agains hydrolysis are highly related to pH. However, very surprisingly, not yet found a paper giving a table/chart of their...
25 July 2024 8,738 0 View
In running two-dimensional gel electrophoresis on bacterial protein, some spots that appear to match a protein sequence have a significantly more acidic isoelectric point than the calculated pI....
24 July 2024 8,076 3 View
I found that several expression constructs of published protein structures containing an N-teiminal fusion tag GAMGSGIQRPTST. What's the founction of this fusion tag?
23 July 2024 1,353 4 View