Streptavidin Magnetic Beads for DNA-protein pull-down assay

Overview

Tested applications

For coupling of DNA Oligonucleotide. Optimal amount should be determined by the end user.

Product Picture

Specificity

Binding to biotinylated molecules, e.g. DNA Oligonucleotide.

Properties

Form

Beads in Suspension (10 mg/mL), supplied in phosphate buffered saline (PBS) pH 7.4, with 0.1% bovine serum albumin (BSA) and 0.02% sodium azide as preservatives.

Storage instruction

Store at 4°C. This product is stable for 12 months. Do not freeze, dry or centrifuge beads. This may cause irreversible aggregation and decreased binding capacity.

Bead Diameter:	1 µm
Concentration:	10 mg/mL
Binding Capacity:	>100nmol Free biotin/mL beads slurry

 

Applications

Application Image

Figure 1. DNA Oligonucleotide pull-down assay for promoter binding transcription factors. A lysate is made of isolated nuclei from bone marrow-derived macrophages (here the macrophages were activated with the cytokine interferon gamma (IFNg)). To this lysate, the biotinylated custom oligo is added. The 20mer oligo corresponds to the promoter of a gene of interest; in this case the gene was inducible nitric oxide synthase (iNOS). Transcription factors bind the biotinylated oligo and these protein:DNA complexes are then isolated using streptavidin magnetic beads. The complexes are then subjected to SDS-PAGE and Western blotting, in this example, for the transcription factor STAT1. STAT1 translocates to the nucleus with interferon gamma treatment (top panel (+)) and is phosphorylated/activated (middle panel (+)). The pull down panel (bottom) demonstrates that in response to IFNg treatment STAT1 binds to the iNOS promoter.