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Questions related from Sinem Ulusan
Hello, I am trying to clone, express and purify a mammalian protein (enzyme) from E.coli BL21 using pet28 expression vector. The vector has N terminal His tag however, because of some cloning...
06 August 2019 7,225 10 View
Hello all, I have a C-terminal his-tagged protein around 70kDa that I want to purify using IMAC technique. I have used both Ni-NTA and Co-NTA columns from different companies and I have used both...
01 July 2019 6,976 16 View
Hello everyone, I am having a trouble in keeping my plasmid inside the bacteria. My plasmid is a Lentiviral plasmid of 7 kb and with the insert that I cloned is now 9 kb. The resistance gene is...
21 October 2018 9,335 4 View
Hello, I am trying to purify a mammalian enzyme from bacteria E.coli (BL21(DE3)) and I have cloned the gene into pet28a and start to do induction with IPTG. After lysis with Lysozyme containing...
16 February 2018 679 6 View
Hello, I am trying to induce my BL21 cells with IPTG and I will purify my his tagged- protein using Ni-NTA column. I am using a lysis buffer of 25mM Tris-Cl, 50mM glucose and 10mM EDTA pH8.0 and...
13 December 2017 7,461 4 View
Hello, I am working with an enzyme kinetic activity in which the substrate is a fatty acid.. After I did my measurements I was cleaning the plate with deionized water but when I re-use it I am...
18 August 2017 8,187 5 View
Hello, I am having trouble in preparing some standard solutions for my enzyme of interest to check its enzymatic activity. In the product sheet it says it contains > 50000 units/mg of solid and...
12 June 2017 1,991 7 View
