Hefin Jones

6 Questions 15 Answers 0 Followers

Questions related from Hefin Jones

How long can I incubate fibroblasts with golgi block?

Hi all, I'm working with primary human synovial fibroblasts in culture. I have a couple of plasma membrane proteins which are upregulated nicely after 24 hours of treatment (quantified by...

15 September 2015 2,610 6 View

Why would macrophages round up and detach after an hour in PBS?

Hi, we have a protocol from a manufacturer of a Ca2+ flux assay which requires macrophages be incubated with the dye (FURA-3) in sterile PBS without Ca2+ or Mg2+ for 1 hour at 37C. We noticed that...

02 July 2015 9,969 4 View

What are the most reliable phenotypic markers for fibroblasts?

Hi! I don't work with fibroblasts usually but with macrophages and could use some advice. I want to do some transwell co-culture experiments with macrophages and fibroblasts (primary, isolated...

12 May 2015 3,453 4 View

Why would macrophages upregulate CD86 and MHCII in response to IL-4?

Hi all, I'm working with primary, human monocyte-derived macrophages and have been polarizing them in vitro with LPS+IFNy or IL-4 (or medium alone). I've done this many times before in human and...

26 March 2015 571 9 View

Why would cells detach with FURA-2 staining?

Hi, We've been trying to perform Ca2+ flux experiments by dying cells with FURA-2, and then measuring fluorescence using a plate reader in a 96-well format. The protocol has worked beautifully...

26 March 2015 9,379 3 View

How can I calculate the sedimentation time of vesicles?

Hi all, I'm trying to calculate the theoretical time for a vesicle to sediment during centrifugation. I have (I think) all the information, but the numbers I'm getting out don't seem to make...

04 March 2015 7,533 3 View