Samuel Davis

6 Questions 55 Answers 0 Followers

Questions related from Samuel Davis

Improving TEV cleavage efficiency - does 50 mM biotin affect activity?

Hi, I'm expressing and purifying a membrane protein which at the C-terminus has a short linker followed by a TEV-mEmerald-TwinStrep tag. I can solubilise and purify the protein with...

02 January 2021 2,475 7 View

Dissolving Cholesteryl hemisuccinate tris salt in detergents?

Hi, Just wondering if anyone has used the Cholesteryl hemisuccinte (CHS) tris salt before, more specifically with different detergents for membrane protein solubilisation? I have previously...

12 December 2019 2,689 10 View

Lipid supplementation of insect cells - any experience?

Hi, I'm attempting to use insect cells (SF9 or High Five) to express a pentameric ligand gated ion channel membrane protein, similar ones of which have historically not been very successful in...

16 October 2018 5,793 3 View

Culturing insect cells in Graces Insect Media - is CO2 required?

Hi, For insect cells (SF9/High Five) we normally culture in SF900-III or Express Five serum free media, but I am looking at using cells cultured in Graces media for a specific experiment. I'm a...

16 October 2018 4,172 0 View

In-gel GFP fluorescence with SDS-PAGE protocol?

Can anyone give me some pointers on how to carry out an SDS-PAGE in-gel fluorescence assay with a uvGFP tagged membrane protein? From various protocols I've come across, I understand that I can...

09 September 2018 6,783 9 View

Problems removing RT- bands in RT-PCR - any suggestions?

Hi, At the moment I'm trying to acertain the endogenous gene expression of hP2Y receptor subtypes in the tsA201 cell line by reverse transcriptase PCR, however I keep having problems with bands...

29 June 2016 7,611 13 View