Kits
The assay kit is designed to measure B-Raf dependent phosphotransferase activity in a kinase cascade reaction using recombinant MEK1, unactive as a B-Raf substrate. Recombinant MAPK2/Erk2, unactive is phosphorylated and activated by the activated MEK1 leading to phosphorylation of a MAP Kinase substrate, myelin basic protein (MBP). [γ-32P]ATP is the final phosphate donor.
Intended for the assay of recombinant B-Raf (supplied) kinase activity with experimental variables as determined by the user. For assay of B-Raf activity from a cell lysate, we recommend an immunoprecipitation-kinase assay, using anti-B-Raf antibody (07-453).
Intended for the assay of recombinant Raf-1 (supplied) kinase activity, with experimental variables as determined by the user. For assay of Raf-1 activity from a cell lysate, we recommend an immunoprecipitation-kinase assay, using anti-Raf-1 antibody (07-396).
Chromatin Immunoprecipitation (ChIP) is an important technique allowing the researcher to analyze in vivo interactions of proteins with genomic DNA. Any chromatin-associated or DNA binding protein can be analyzed with this technique, provided a good antibody to the protein exists. One can measure different proteins localized to a specific region of the genome, or the genome wide distribution of a specific protein. Another powe…
The 384 well format kit is a newer version of the tried and true non-radiographic HDAC assay. As in the previous kit, samples are first incubated with the HDAC Assay substrate, allowing deacetylation of the fluorometric substrate. After incubation, the activator solution is added, and releases a fluorophore from the deacetylated substrate or standard. The sample is then put into a microplate reader and fluorescence (i.e free/c…
This assay is a simple two-step procedure performed in a microtiter plate. In the first step, samples are incubated with the HDAC assay substrate, allowing deacetylation of the substrate. Next, the Activator Solution releases p-nitroanilide from the deacetylated substrate or standard.
The EZ-Zyme™ Chromatin Prep kit allows ChIP analysis at nucleosome resolution by performing complete or partial digestions with a proprietary enzymatic cocktail to obtain chromatin fragments of on average one to a few nucleosomes in length. Compared to sonication, this procedure not only allows mapping the protein-DNA association at a higher resolution, but also renders the subsequent immunoprecipitation more efficient due to…
The Rac1/Cdc42 Activation Assay provides an effective method for detecting Rac and Cdc42 activity in cell lysates. This assay uses the downstream effector of Rac/Cdc42, p21-activated protein kinase (PAK1), to isolated the active GTP-bound form of Rac/Cdc42 from the sample. The p21 binding domain (PBD) of PAK1 is expressed as a GST-fusion protein and coupled to agarose beads. After precipitation, an immunoblot is performed and…
The ChIPAb+ CREB, antibody/primer set includes the CREB, antibody, a negative control antibody (purified rabbit IgG), and qPCR primers which amplify a 64 bp region flanking the CRE of the human cFos gene. The CREB and negative control cultured supernatant are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of CREB associated chromatin.
Every lot of the ChIPAb+ line of antibodies is individually validated for chromatin precipitation, in order to guarantee successful ChIP assays every time. Each antibody includes a control primer set for performance confirmation. Sp1 antibody and the negative control antibody (rabbit normal IgG) can be used to demonstrate that the Sp1 antibody is functionally validated in the precipitation of Sp1 associated chromatin. The qPCR…
