20158-LightShift-RNA-EMSA-250

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 KitLightShift Chemilumin-
escent RNA EMSA
RNA Protein Pull-Down Kit
TargetProtein-DNA interactionProtein-DNA interactionProtein-DNA interactionsProtein-RNA interactionProtein-RNA interaction
Interaction conditionsIn vitroIn vivoIn vivoIn vitroIn vitro
Nucleic acid labeling methodBiotinNANABiotinDesthiobiotin (included in kit)
Base beadNADynabeads Protein A/G Magnetic beadsChIP-grade Protein A/G Plus AgaroseNAPierce Nucleic Acid-Compatible Streptavidin Magnetic Beads
Primary advantageNon-radioactive detectionQuantitativeQuantitativeNon-radioactive detectionEnrichment of low-abundance targets
Detection methodWestern blotqPCRqPCRWestern blotWestern blot or mass spectrometry
Preparation and sample processing time4.5–5 hrs5.5 hrs7.5 hrs<8 hrs3 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.

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.

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.

For Research Use Only. Not for use in diagnostic procedures.