Regan Miller

6 Questions 10 Answers 0 Followers

Questions related from Regan Miller

Protocol for primary AML cell co-cultures?

I'm curious to know if anyone has a cell culture protocol for a co-culture of AML primary samples with either stromal or endothelial cells? I have very limited samples so no room for "trial and...

16 October 2019 6,474 0 View

Cloudy media that doesn't seem to affect cell growth?

I've been working with a primary endothelial cell line given to us by a collaborator. These cells can only be passaged once, so I grew them to around 70% confluency in a T-75 with no issues. I...

23 August 2018 276 2 View

Cell pellet after addition of RIPA?

I've been performing protein extractions/Bradford assays no problem. Then the other day one of my cell pellets would not dissolve in RIPA/Halt protease inhibitor, leading to extremely low...

06 February 2018 9,121 2 View

Reasons for blurry background on blot other than overloading protein?

I ran a Western earlier in the week and when I read my result, there were ghost bands, non-specific bands, and a dark background. At first we thought I loaded too much protein (40 ug on a 0.45...

19 January 2018 2,156 6 View

Why would there be differences between 0.2 um and 0.45 um pvdf membranes?

Relating to my recent question about high background, I ran a new test. I took un-probed pvdf (both 0.2 um and 0.45 um) and activated them in methanol, put them in water for a few minutes, blocked...

28 September 2017 8,966 1 View

What are some reasons for having an extremely dark background on a pvdf membrane after reading on chemidoc?

I'm relatively new to Western blotting and I've been recently having issues with an extremely dark background on my pvdf membrane. This time I paid close attention to wetting it in methanol (what...

18 September 2017 7,518 5 View