Umar Burki

6 Questions 2 Answers 0 Followers

Questions related from Umar Burki

Is there a way to encapsulate cell membranes in order to prevent intracellular oligos and proteins from leaking?

Aim: I am isolating adult cardiomyocytes from mice injected by our therapeutic oligo and my aim is to measure its uptake. Method: I am able to isolate viable cardiomyocytes using the classic...

02 November 2017 8,998 1 View

How to remove satellite cells from isolated mouse muscle fibres?

I need to quantify the uptake of our therapeutic oligo drugs in the nuclei of muscle fibres isolated from adult mice. However these fibres have several satellite cells attached to them (see photo...

10 January 2017 9,660 3 View

Has anyone used Fluorescence in situ Hybridization (FISH) for detecting oligonucleotides in mouse tissues or cells?

I am trying to develop a Fluorescence in situ Hybridization (FISH) assay for visualising the sub-cellular localisation of 25-mer oligonucleotide drugs in mouse tissues. The aim is to show the...

05 February 2016 288 5 View

Is 20mM of extracellular calcium concentration toxic to cells in vitro?

During Fluo-4 calcium influx assay in human fibroblasts, I find that adding 2mM Ca in Ca-free well doesn't increase fluorescence intensity (FI) even over 10min period, which is expected. However,...

19 November 2015 8,474 5 View

Why does addition of water alone increases fluorescence of Ca influx in fibroblasts loaded with Fluo-4 NW dye?

I am using a Tecan plate reader to develop a high throughput fluorescent Ca imaging assay for measuring Thapsigargin stimulated calcium influx in human fibroblasts. The cells are seeded in clear...

24 June 2015 1,914 2 View

Why are single muscle fibers from mice hyper-contracting during isolation?

I'm trying to isolate single muscle fibers from healthy C57/BL10 mice for downstream assays such as intracellular calcium, sodium and pH measurements. I'm isolating the fibers from EDL...

02 April 2015 9,426 3 View