Cell Signaling Technology

Product Pathways - Chromatin Regulation

ChIP-Grade Protein G Agarose Beads #9007

Description

ChiP-Grade Protein G Agarose Beads are an affinity matrix for the small-scale isolation of immunocomplexes from chromatin immunoprecipitations (ChIP assays). A truncated form of recombinant Protein G is covalently coupled to agarose beads. Protein G exhibits high affinity for subclasses of IgG from many species (including human, rabbit, mouse, rat and sheep) and can be used for immunoprecipitation assays with these antibodies. The beads are stored in buffer containing BSA (500 µg/ml) and sonicated salmon sperm DNA (200 µg/ml) to block non-specific binding of proteins and DNA during isolation of immunocomplexes.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed using digested chromatin from HeLa cells and the indicated antibodies. Purified DNA was analyzed by quantitative Real-Time PCR, using primers specific for the transcriptionally active RPL30 gene, the inactive MYOD gene or the heterochromatic α satellite repeat element. The relative abundance of each DNA sequence enriched by protein-specific immunoprecipitations is compared to the amount of the same DNA sequence enriched by the non-specific Normal Rabbit IgG #2729 (background).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed using digested chromatin from HeLa cells and either Histone H3 Antibody #2650 (lane 2), Rpb1 CTD (4H8) Mouse mAb #2629 (lane 3), Di-Methyl Histone H3 (Lys9) Antibody #9753 (lane 4), or Normal Rabbit IgG #2729 (lane 5). Purified DNA was analyzed by standard PCR methods using primers specific for the transcriptionally active RPL30 gene, the inactive MYOD gene or the heterochromatic α satellite repeat element. PCR products were observed for each primer set in the input sample (lane 1) and various protein-specific immunoprecipitations, but not in the immunoprecipitation using Normal Rabbit IgG #2729 (lane 5).

Directions for Use

Vortex tube briefly to resuspend the beads. Add 30 μl of bead slurry to each chromatin immunoprecipitation (ChIP) reaction. For bead washing and subsequent elution of immunocomplexes, the beads can be separated from solution by a brief 1 minute centrifugation in a microcentrifuge at 6,000 rpm. Resuspend the beads in solution by gently vortexing or rocking the tube.

Storage

Supplied in 10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 500 µg/ml BSA, 200 µg/ml sonicated salmon sperm DNA and 0.05% sodium azide. Store at 4°C.

Application References

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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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