14 Questions 8 Answers 0 Followers
Questions related from Tara Stirland
Hi I'm looking for some help in analysing my luciferase data. I have a transcription factor and then a range of proteins I think will influence transcription factor activity. For each...
09 November 2018 4,794 3 View
Hi I am looking for protein protein inteactions between a shuttle protein and nuclear hormone receptor using BIFC. When I perform BIFC with Fos and Jun I get bright signal and when I use Mutated...
22 February 2018 2,067 1 View
Hi I have performed a gene card with 96 genes to see whether certain genes are expressed or not between skin models and full thickness skin. In some cases, genes are not expressed and so I get an...
03 July 2017 8,767 6 View
Hi I'm looking at cloning into pINDUCER20. Having only ever cloned into pcDNA3.1 and pet28A before, I'm unsure of where to start. I think I need to do recombination but don't really know much...
27 March 2017 7,004 0 View
Hi I'm growing HaCaT cells and since taking this batch out of liquid nitrogen I've started to observe bubble or vacuole like structures in approx. 5-10% of cells. I've attached a representative...
27 February 2017 8,413 2 View
Hi I am looking at producing lipid vesicles to use in an experiment. The paper I am following talks about mixing lipids 40 mol % and 60 mol % to total of 50 uM phospholipids. Does anyone know what...
07 December 2016 4,336 10 View
Hi I usually work with primary keratinocytes but am looking at performing some work with HaCaT cells - can I ask what is the optimal passage to use these up until? I currently am growing some at...
06 December 2016 4,745 1 View
Hi everyone I am try to transfect primary human keratinocytes for downstream use in a luciferase reaction. Thus far I have been trialling X-tremeGENE HP in a 1:3 and 1:2 ratio on keratinocytes...
03 June 2016 2,075 3 View
Hi I am try to show that an organotypic skin model I have built closely resembles the structure of human skin by performing immunofluorescence with antibodies directed against a number of antigens...
23 May 2016 5,385 2 View
Hi, I am looking to prepare 5637 conditioned media for use in Mutz3 media and was wondering if anyone had experience with this? Many thanks
26 January 2016 2,375 1 View
Hi, I am looking at building a 3D skin model and wish to follow a protocol using a 6 well plate insert but I have 24 well plate inserts. What would the scale factor be? Many thanks
03 November 2015 3,674 3 View
Hi, I am trying to design a custom TaqMan® Gene Expression Assay for the first time and am having issues with some of the genes that I wish to include. In some cases, it tells me that the best...
09 August 2015 998 2 View
Hi, I am currently isolating keratinocytes from human skin samples but I am having trouble with the speed of keratinocyte growth. I am using overnight treatment in 5mg/ml dispase to disociate the...
07 May 2015 7,883 4 View
Hi, I am isolating keratinocytes, melanocytes and fibroblasts from human skin and wish to use immunocytochemistry to confirm that the cells are the type I say that they are. Has anybody got any...
03 May 2015 6,446 3 View
