Gregory James Buchan

9 Questions 21 Answers 0 Followers

Questions related from Gregory James Buchan

Would dimethyl fumarate work for critically-ill Covid19 patients?

Hello, Dimethyl fumarate is a readily-available, FDA-approved drug for multiple sclerosis. It can both inhibit NF-kB and activate Nrf2. This could potentially limit the severity of viral-induced...

31 March 2020 3,311 7 View

How can we compare DC studies that use different methods to isolate them?

Hello, I've been (not so successfully) trying to perform basic experiments on dendritic cells to analyze the actions of a fatty acid. I've noticed there are numerous methods to do this and it...

06 February 2019 9,627 6 View

Is b-mercaptoethanol (BME) required for immune cells culture?

Hello, Many protocols add b-mercaptoethanol to cell media (concentrations around 0.05 mM) in order to keep T cells, macrophages, and DC cultures viable; however, some protocols do not. BME is a...

24 September 2018 7,723 5 View

How do I culture more CD11c+ BMDC from mice?

Hello, I am new to harvesting BMDC and my most recent experiment had 45% CD11c+ cells that were viable (also many dead cells, only 20-30% viable). I followed the method from Madaan A, et al....

19 July 2018 5,833 3 View

How much EDTA should be used for flow cytometry purposes?

Hello, I've been perusing the literature and a variety of protocols use EDTA concentrations from 0.03-5 mM EDTA in their buffers for isolating immune cells from the lung and/or BALF for flow...

22 January 2018 2,606 3 View

How do you use cell lysates for studying enzyme kinetics?

Hello, I'm trying to determined the Vmax, Km, etc...for a membrane bound short chain dehydrogenase. I am planning on using cell lysates as a source of the enzyme. Do you know of any protocol to...

07 April 2016 1,608 4 View

Are there standard media conditions for treating influenza in vitro?

Hello, I am trying to treat A549 cells infected with PR8 and I am trying to determine the best media conditions but in the literature conditions are all over the place. I see 2% FBS, 5% FBS, 10%...

28 January 2016 2,028 4 View

Why aren't A549 lung cells used for influenza plaque assay?

I am trying to determine viral titer and we do not have MDCK cells. My logic tells me influenza replicates in lung cells and not kidney cells and yet MDCK has been established for many years. Why...

24 January 2015 4,189 6 View

How does reaction time effect ligation?

Typical ligation protocols (for inserting a certain gene/deletion into a plasmid) recommend a ligation of 1-24 hrs. Has anyone seen significant differences in yield between 1 hr vs. 24 hr? Thanks

06 September 2014 2,841 3 View