Timothy Keiffer

4 Questions 3 Answers 0 Followers

Questions related from Timothy Keiffer

How can I troubleshoot this STAT1 homodimerization assay?

I am trying to detect STAT1 homodimerization by first transfecting 293T cells with 2 STAT1 constructs, one with a GFP tag and the other with a FLAG tag, and then treating cells with either...

09 September 2018 4,627 0 View

What effects do urea and guanidinium chloride have on vesicles?

If I'm trying to isolate a vesicle without detergent, could I possibly use urea and/or guanidinium chloride to facilitate detachment of the vesicle into the cytosolic fraction of cells? Or does...

05 May 2015 4,401 1 View

Any way to detach bound proteins from microtubules in vitro from a nuclear extract?

Is there a way to detach proteins from microtubules (spindle microtubules) from a nuclear extract in vitro? I've tried to do this in vivo by treating living cells with nocodazole before...

05 May 2015 246 4 View

Any optimization suggestions or information blunt-end cloning?

Here is my dilemma: I am trying to insert a 3.7kb sequence into the plasmid pMA3211 (for tet-inducible expression of insert sequence), which is 6.7kb. First, I digested pMA3211with SmaI (blunt)...

04 April 2014 6,050 4 View