study both transcriptional & post-transcriptional regulation
Though gene regulation studies have historically focused on transcription, gene expression is controlled at both the transcriptional and post-transcriptional levels. Transcription factors and other DNA-binding proteins modulate when, where and how much of a gene’s mRNA is transcribed, while microRNAs and the RISC complex interact with the 3´UTR of gene transcripts to control the stability of an mRNA and its translation into protein. In addition, because the DNA is packaged as nucleosomes, histone modifications and DNA methylation play a large regulatory role by altering chromatin structure, which influences whether or not the DNA is accessible for transcription.
Active Motif offers a variety of products for studying each of these types of gene regulation. Brief summaries for some of these products are shown below, with links to individual product pages for more information.
Products for the study of Transcriptional Regulation
- TransAM® Transcription Factor Assays are DNA-binding ELISAs that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. With assays available for over 40 different targets and over 800 papers citing their use, TransAM Kits have become the method of choice for detecting and quantifying activated transcription factors.
- The LightSwitch™ Luciferase Assay System includes over 18,000 promoters that have been cloned from the human genome into transfection-ready LightSwitch reporter vectors, which are assayed in living cells using our optimized luciferase assay reagents and protocols. Also available are Synthetic Response Element constructs and Pathway Reporter Stable Cell Lines.
- Luminex® Transcription Factor Multiplex Kits are novel bead-based assays that enable simultaneous, comparative analysis of multiple members of the NFκB or AP-1 transcription factor in the same sample.
- FunctionELISA™ IκBα is a sensitive way to monitor the phosphorylation of IκBα, a protein that is highly studied due to its important regulatory role in inhibiting the activation and translocation of NFκB dimers.
- Our Nuclear Extract Kit simplifies the task of isolating high-quality nuclear, cytoplasmic or whole-cell extracts from mammalian cell or tissue samples. We also offer over 100 ready-to-use Cell & Tissue Extracts, many prepared following stimulation to activate specific transcription factors.
- Co-Immunoprecipitation is often used to study protein/protein interactions. Our Nuclear Complex Co-IP Kit has been optimized for use on nuclear complexes, while our Universal Magnetic Co-IP Kit utilizes magnetic beads for Co-IP of either nuclear or cytoplasmic protein complexes.
- The electrophoretic mobility shift assay (EMSA), also known as gel shift, has long been used to study protein-DNA binding events. Our Gelshift™ Chemiluminescent EMSA Assay Kit greatly improves on the technique by eliminating the use of radioactivity through use of a more sensitive, chemiluminescent detection method.
- Nuclear receptors (NR) belong to a superfamily of structurally related ligand-inducible transcription factors. Our NR Sandwich ELISAs make it simpler to study the Androgen, Estrogen and Progesterone receptors.
Products for the study of Post-transcriptional Regulation
- The LightSwitch™ 3´UTR Luciferase Reporter Collection includes over 12,000 3´UTRs cloned from the human genome and supplied as transfection-ready LightSwitch reporter vectors. Used with our miRNA Mimics and Inhibitors, you can validate miRNA targets as well as measure RNA stability, translation efficiency and the functional impact of miRNAs on a gene-by-gene basis.
- In addition to our endogenous human 3´UTR clone collection (see previous bullet), we offer over 900 LightSwitch™ Synthetic miRNA Target Reporter constructs; these contain optimized sequence repeats that can provide better sensitivity than their native 3´UTR counterparts because they are fully complementary to human and human viral miRNA sequences.
Products for the study of Chromatin, Histones & Nucleosomes
- Chromatin Immunoprecipitation (ChIP) is used to map epigenetic marks and the binding of transcription factors and other DNA-binding proteins to specific loci in the genome. Active Motif's ChIP-IT® line of kits are the most complete available, and includes specialty kits specifically optimized for a variety of ChIP procedures such as ChIP-Seq, sequential ChIP, RNA ChIP, High Sensitivity ChIP and ChIP from FFPE samples.
- Chromatin is subject to a variety of chemical modifications that directly influence gene regulation, including post-translational modifications of the histone tails and methylation of cytosine residues in the DNA. Active Motif offers comprehensive product lines for the study of Histones and Histone Modifications as well as DNA Methylation.
- The position of a nucleosome at a gene's promoter region has been shown to play a large role in transcriptional regulation. Such nucleosomes must be unfolded or disassembled in order for initiation of transcription to occur. The NOMe-Seq Kit is the first assay able to provide both a nucleosome footprint and a DNA methylation profile for the same DNA strand. This enables researchers to study the role of nucleosome occupancy in the context of DNA methylation for their target gene of interest.