15 Questions 8 Answers 0 Followers
Questions related from David J Walker
I'm planning to make conditioned media from various cell lines for some experiments and have come across various protocols, which all differ in their methods. My current plan is: 1) Culture...
12 January 2023 8,340 2 View
I've been piloting clonogenic survival assays to look at long-term effects of of drugs in colon cancer cell lines (SW620, SW480, HT-29). My initial pilot assays in 12-well plates were successful...
18 March 2022 7,567 1 View
I've been trying for several weeks to get consistent and successful transient transfection of glosensor DNA (promega) into Sw620 and Sw480 colon cancer cells in order to measure cAMP levels by...
23 November 2021 8,182 3 View
Hi there, I've developed an algorithm in FIJI for quantifying Iba+ cells via DAB counterstain on IHC brain sections. My algorithm does a pretty good job of identifying the soma which allows for...
29 July 2020 3,438 3 View
I've been working with murine neural stem cells as a 2d monolayer which is achieved by adding laminin (ECM) directly to the culture media, without the need to coat the TC plastics. However, I've...
05 February 2020 8,267 2 View
I have multiple sig pairwise comparisons generated from a 3-way interaction from a mixed model GLM. I want to present these comparisons in my thesis but wish to do it concisely so it is easy to...
15 June 2018 8,065 3 View
Hi there, I need assistance with interpreting the outcome of my pairwise comparisons from my datasets. I've been running a glmer mixed models and selecting the best model using the AIC criteria....
28 July 2017 220 2 View
Hi, I have several dual-labelled fluorescent images of brain tissue where I have obvious colocalisation between my protein of interest (BDNF, imaged at Alexafluor 594) and the nuclei (DAPI...
18 December 2016 9,280 2 View
Hi there, Yet another IF-related question. I'm going to be dual-labelling free-floating tissue sections with fluorescent secondary abs. After the final wash steps, I will be mounting sections...
23 June 2016 7,758 3 View
Hi there, I'm planning some immunofluorescence where I want to dual label either BDNF or TRKB with caspase-3. However, for the species of tissue I'm using, I need to use an antibody that hasn't...
29 May 2016 4,322 2 View
I've come across a neat protocol for measuring immune response in plasma which uses bioluminescence. However, I was wondering if it was possible if the same samples can then be processed for...
04 March 2015 9,938 1 View
I'm looking for an IHC or mRNA marker that specifically identifies ramified (unactivated) microglia so I can distinguish between an inactive and active phenotype. Can anyone help please? Thanks
29 January 2015 9,844 8 View
I am preparing ACSF for organotypic slice culture experiments (which I haven't done before) but I want to make a 10x stock and I want to make sure I have separated the correct components as some...
08 January 2015 2,611 0 View
My question has two parts re the organotypic slice culture technique: How long can tissue (in this case brain) remain embedded in agarose prior to slicing on vibratome (4% low-melt agarose in...
27 December 2014 6,267 1 View
I am looking for papers detailing a method to isolate primary neurons from an avian species and establish them in culture. In particular, I would like an idea of what sort of cytokines/growth...
13 March 2014 8,595 2 View
