Lincoln Shane

6 Questions 5 Answers 0 Followers

Questions related from Lincoln Shane

Is it advisable to clone and express mammalian genes in bacteria?

Hello, For all i know it is well known mammalian proteins undergo extensive PTMs immediately after translation to augment protein function diversity. But expressing these proteins in bacteria -...

01 January 2019 8,216 0 View

Do i need to use coding sequences to clone a functional human gene into bacteria?

Hello colleagues! I have been to trying to express a human gene but after encountering many challenges, i took a pause to revaluate my strategies. For mammalian genes particularly those that code...

01 January 2019 1,545 4 View

Where do i place the codon or do i need it at all?

Two, perhaps as you probably are aware, it is recommended that a stop codon be placed at the end of the insert. But as it turns out, most commercial plasmids come with stop codons at the end of...

01 January 2019 5,319 5 View

What can you tell me about an E.coli strain named Lemo21 from NEB?

I am currently trying to express human tyrosinase in E.coli and as you know this enzyme being a membrane protein it comes with a fair share of difficulties relating to expression in BL21(DE3)....

10 October 2018 7,813 3 View

Protein purification protocol?

Hello fellow scientists, here i have a bit of a seemingly demoralizing issue with protein purification. Could you please help? I have attached a commassie image for clarity. 10mM imidazole, 50 mM...

12 December 2017 3,126 5 View

Hello everyone out there?

I have a question regarding western blotting of large proteins particularly the clostridium difficile toxins. What SDS-PAGE (Stacking and Resolving gel) concentrations should i use? Even so, can i...

09 September 2015 9,037 1 View