Amila Zuko

5 Questions 17 Answers 0 Followers

Questions related from Amila Zuko

How to avoid uneven (intensity) bands during Western Blotting?

I've uneven bands (size, intensity, and staining) when doing Western Blotting and I'm wondering what the problem could be. The protocol: -10% separating gel (0.375M Tris pH8.8; 0.1% SDS; 10%...

07 July 2019 1,863 9 View

How to reduce the amount of coeluates after IMAC?

I'm performing IMAC to purify my his-tagged protein (enzyme) from e. coli lysate. It seems I can get a good amount of my protein of interest but the problem is that I also get alot of coeluates,...

10 October 2016 7,053 7 View

How to dissolve PEI (polyethylenimine)?

This concerns linear PEI 25kDa. So far I am aware of two methods: 1: stir the PEI in MQ (1mg/ml) and dissolve by dropping and holding the pH at 2. After its transparant increase pH to 7 -->...

04 April 2016 1,432 17 View

What could be the reason for these strange chromatograms (see attachment) after DNA sequencing?

Lately I'm getting failed DNA sequences and strange sequence peaks in my chromatograms when trying to sequence my plasmid (see attachments). With a previous mini prep sample, the following...

02 February 2016 3,899 4 View

Could someone help me with some Neuronal Tissue Co-IP issues?

I have been doing co-immunoprecipitations for a while now using my own buffers (CHAPS lysis buffer + protease & phosphatase inhibitors) and Dynabeads protein G beads. Useful information: -The...

01 January 2014 2,360 4 View