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Protein-Nucleic Acid Interactions
We offer a selection of easy-to-use kits for chromatin immunoprecipitation assays (ChIP), electrophoretic mobility shift assays (EMSA), and DNA pull-down assays. These kits allow you to determine which proteins interact with DNA or RNA, and analyze what proteins are present in these protein-nucleic acid complexes. The next step, identifying the nucleic acid sequences required for the assembly of these complexes facilitates the ultimate understanding of the regulation of specific cellular processes.
- More choices—different strategies to capture various protein-nucleic acid interactions and downstream analysis needs
- Complete—include all controls and reagents needed for application
- Fast—all kits are designed to minimize preparation and sample processing time
Which protein-nucleic acid interaction product is right for me?
 |  |  |  |  | |
|---|---|---|---|---|---|
| LightShift Chemilumin- escent DNA EMSA | MAGnify Chromatin Immunopre- cipitation System | Pierce Agarose ChIP Kit | LightShift Chemilumin- escent RNA EMSA | RNA Protein Pull-Down Kit | |
| Target | Protein-DNA interaction | Protein-DNA interaction | Protein-DNA interactions | Protein-RNA interaction | Protein-RNA interaction |
| Interaction conditions | In vitro | In vivo | In vivo | In vitro | In vitro |
| Nucleic acid labeling method | Biotin | NA | NA | Biotin | Desthiobiotin (included in kit) |
| Base bead | NA | Dynabeads Protein A/G Magnetic beads | ChIP-grade Protein A/G Plus Agarose | NA | Pierce Nucleic Acid-Compatible Streptavidin Magnetic Beads |
| Primary advantage | Non-radioactive detection | Quantitative | Quantitative | Non-radioactive detection | Enrichment of low-abundance targets |
| Detection method | Western blot | qPCR | qPCR | Western blot | Western blot or mass spectrometry |
| Preparation and sample processing time | 4.5–5 hrs | 5.5 hrs | 7.5 hrs | <8 hrs | 3 hrs (hands-on time only) |
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Featured data for protein-nucleic acid interaction products
Chemiluminescent EMSA of four different DNA:protein complexes.
Biotin-labeled target duplexes ranged in size from 21-25 bp. The EBNA reactions were supplemented with 2.5% glycerol and 0.05% NP-40, and the AP1 reactions were supplemented with 10% glycerol. The source of the Oct-1, AP1 and NF-κB transcription factors was a HeLa nuclear extract. EBNA-1 extract is provided as a control in the kit. Unlabeled specific competitor sequences (where used) were present at a 200-fold molar excess over labeled target. X-ray film exposure times for each system ranged from 2 minutes for EBNA, Oct-1 and AP1, and 5 minutes for NF-κB. |  |
Thermo Scientific ChIP Pierce Grade Protein A/G Agarose Beads increase specific signal and decrease background signal in the ChIP assay as compared to ChIP-qualified Protein G agarose beads from other leading suppliers.
Following formaldehyde crosslinking, HeLa cell lysate was prepared using the Chromatin Prep Module of the Pierce ChIP Assay Kit. Immunoprecipitations were performed using an anti-RNA Polymerase II (RNA Pol II) antibody or normal rabbit IgG (IgG). Antibody-antigen complexes were then recovered with blocked or unblocked Pierce Protein A/G Plus Agarose Beads or ChIP-qualified Protein G agarose beads from two independent suppliers. All other steps in the ChIP assay were performed using the Pierce ChIP Assay Kit. Quantitative PCR was performed using primers that amplify the proximal promoter region of the human GAPDH promoter.
Example RNA-protein pull-down experiment: the effect of mutation on the ability of androgen receptor 3′-UTR RNA to bind HuR and Poly(C)BP.
Wild-type and mutant AR 3′-UTR RNA (50 pmol) were labeled and used for the pull-down assay according to the kit procedure. Samples were normalized by volume.L = lysate load; FT = flow-through, E = eluate. PCBP1 antibody (1:1000).
Chemiluminescent EMSA of four different DNA:protein complexes.
Biotin-labeled target duplexes ranged in size from 21-25 bp. The EBNA reactions were supplemented with 2.5% glycerol and 0.05% NP-40, and the AP1 reactions were supplemented with 10% glycerol. The source of the Oct-1, AP1 and NF-κB transcription factors was a HeLa nuclear extract. EBNA-1 extract is provided as a control in the kit. Unlabeled specific competitor sequences (where used) were present at a 200-fold molar excess over labeled target. X-ray film exposure times for each system ranged from 2 minutes for EBNA, Oct-1 and AP1, and 5 minutes for NF-κB. | |
Thermo Scientific ChIP Pierce Grade Protein A/G Agarose Beads increase specific signal and decrease background signal in the ChIP assay as compared to ChIP-qualified Protein G agarose beads from other leading suppliers.
Following formaldehyde crosslinking, HeLa cell lysate was prepared using the Chromatin Prep Module of the Pierce ChIP Assay Kit. Immunoprecipitations were performed using an anti-RNA Polymerase II (RNA Pol II) antibody or normal rabbit IgG (IgG). Antibody-antigen complexes were then recovered with blocked or unblocked Pierce Protein A/G Plus Agarose Beads or ChIP-qualified Protein G agarose beads from two independent suppliers. All other steps in the ChIP assay were performed using the Pierce ChIP Assay Kit. Quantitative PCR was performed using primers that amplify the proximal promoter region of the human GAPDH promoter.
Example RNA-protein pull-down experiment: the effect of mutation on the ability of androgen receptor 3′-UTR RNA to bind HuR and Poly(C)BP.
Wild-type and mutant AR 3′-UTR RNA (50 pmol) were labeled and used for the pull-down assay according to the kit procedure. Samples were normalized by volume.L = lysate load; FT = flow-through, E = eluate. PCBP1 antibody (1:1000).
Related products
Thermo Scientific offers complete and easy-to-use for non-radioactive labeling of DNA and RNA probes in 0.5–2 hr, as well as other reagents for isolating, preparing, and detecting protein-nucleic acid interactions.
Resources
Application notes, tech tips, etc.
For Research Use Only. Not for use in diagnostic procedures.