Microbiology, Bacteria, E.coli
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/General_Information/vol2-issue2-ecoli.pdf
Article A Highly Efficient Procedure for the Extraction of Soluble P...
Article Preparation of Soluble Proteins from Escherichia coli
http://www.epibio.com/docs/default-source/protocols/easylyse-bacterial-protein-extraction-solution.pdf?sfvrsn=8
You can use lysozyme and protease inhibitor cocktail for lysis
http://web.mit.edu/king-lab/www/cookbook/plysis.htm
alternative is to sonicate but be sure to give long pause between on off cycle and between cycles to prevent heating. We generally use 3 cycles of 2s on 5 s off for 2 min and place on ice between cycles for few mins
02 March 2021 3,060 3 View
Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...
02 March 2021 3,584 1 View
Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...
01 March 2021 3,867 3 View
We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...
01 March 2021 3,355 3 View
I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....
01 March 2021 3,607 3 View
Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...
01 March 2021 8,169 2 View
I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...
28 February 2021 7,127 2 View
Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.
28 February 2021 7,883 3 View
28 February 2021 9,936 2 View
I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...
25 February 2021 3,030 1 View