16 Questions 9 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 4,877 3 View
Hi all, I am looking at performing mass-spec of human skin tissue to look at the presence of different phospholipids. Can anyone shed any light on the best protocol to use for this? Many thanks
08 May 2018 6,063 5 View
Hi I am treating CV1 cells with lipid vesicles, which involves a number of washes and an incubation at 2 degrees for 45 minutes. This has lead to extensive but inconsistent cell loss in both...
23 April 2018 7,300 0 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 2,114 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 8,829 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 7,050 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 8,465 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 4,389 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 4,807 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 2,129 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 5,436 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 2,433 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 3,722 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 1,040 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 7,933 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 6,503 3 View
