Open Access Highly Accessed Methodology article

Optimal use of tandem biotin and V5 tags in ChIP assays

Katarzyna E Kolodziej12, Farzin Pourfarzad1, Ernie de Boer1, Sanja Krpic1, Frank Grosveld1* and John Strouboulis13*

Author Affiliations

1 Department of Cell Biology, Erasmus MC, Dr Molewaterplein 50, 3015GE Rotterdam, the Netherlands

2 Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the Netherlands

3 Institute of Molecular Oncology, Biomedical Sciences Research Center Alexander Fleming, PO Box 74145, 16602 Varkiza, Greece

For all author emails, please log on.

BMC Molecular Biology 2009, 10:6  doi:10.1186/1471-2199-10-6

Published: 5 February 2009

Additional files

Additional file 1:

GATA-1 ChIP of the myb promoter. A) Location of ChIP primers in the myb promoter. B) Comparison of different derivatives of immobilized streptavidin: NeutrAvidin, streptavidin agarose, streptavidin mutein and M280 Dynabeads. Relative enrichment is calculated over non-transfected BirA control cells. C) The effects of preclearing chromatin and using 1% FGEL in blocking the beads. Enrichment was calculated relative to negative primer set. D) The effect of omitting SDS from the sonication buffer. Relative enrichment is calculated over non-transfected BirA control cells.

Format: PDF Size: 342KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 2:

GATA-1 ChIP of the GATA-2 gene locus. A) Location of the ChIP primers in regulatory elements of the GATA-2 locus. B) GATA-1 binding using streptavidin M280 Dynabeads. C) The effect of blocking M280 Dynabeads with 1% FGEL. Enrichment was calculated relative to non-transfected BirA cells. Primer sequences are as published by Rodriguez et al. (ref. [8]).

Format: PDF Size: 349KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data