Hi,
I am interested in recommendations of expression vectors for expression of proteins in E.coli and other Gram negative bacteria (potentially) that are independent of the T7 TNA polymerase; eg T5, pBAD, Rhamnose and other.
Does anyone know if sodium azide is cytotoxic to mammalian cells (293, CHO, murine hybridomas) at 0.1-0.2% w/v? Buffers such as MACS buffer seem to include it but I would have thought this would...
03 April 2018 5,669 3 View
Any thoughts on what is the preferred method for glucose feed control in E.coli fed-batch? Both seem to have their merits, but quite difficult to tune pump PID settings to avoid sub-optimally...
31 December 2017 2,608 2 View
Can anyone tell me where i can source this strain? It is a tolRA knockout; hypervesiculating. I cannot find it in the normal collections and correspondence with two corresponding authors who have...
07 August 2017 8,792 1 View
Looking for a good quality antibody to probe tagged proteins produce in E.coli. My last experience with several antibodies showed very different performance from different vendors.
10 November 2015 6,600 4 View
The antibody in question is "humanized" and described in a paper presenting clinical trial data
04 May 2015 5,467 0 View
Can anyone point me to some useful references on methods to assist in clarification of crude homogenates of animal tissue? Centrifugation in our case is only partly successful and our current...
03 April 2014 1,928 1 View
We have purified some of Fabs and would like to get some feedback on the best way to store them. Ideas on • Which buffer? • With/without glycerol? • 4oC or -20 oC? Would be greatly appreciated.
11 December 2013 3,001 2 View
Any suggestions of culture collections or other sources where we could obtain this culture?
09 October 2013 2,336 2 View
I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...
07 August 2024 6,034 3 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
There are anaerobic gas pak jars and environment providers in the laboratory that provide anaerobic environmental conditions for the growth of anaerobic bacteria. However, there is no anaerobic...
05 August 2024 4,945 3 View
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 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
I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...
28 July 2024 7,559 1 View
List down the various factors responsible and how does this effect the overall biogas production when it is related to Hydraulic retention time of the digester based on the feedstock?
27 July 2024 4,800 4 View