Enzyme Activity Assays
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).
The 384 well format kit is a newer version of the tried and true non-radiographic HDAC assay.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 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…
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. Chemicon's Caspase-3 Colorimetric Activity Assay Kits provide a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate DEVD-pNA. The free pNA can be quantified using a s…
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. Chemicon's Caspase-8 Colorimetric Activity Assay Kits provide a simple and convenient means for assaying the activity of caspases that recognize the sequence IETD. The assay is based on spectophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate IETD-pNA. The free pNA can be quantified using a s…
Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. Chemicon's Caspase-9 Colorimetric Activity Assay Kits provide a simple and convenient means for assaying the activity of caspases that recognize the LEHD. The assay is based on spectophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate LEHD-pNA. The free pNA can be quantified using a spectropho…
The CHEMICON Proteasome Activity Assay Kit provides a quick, efficient and sensitive system for evaluation of proteasome activity in cell lysates or inhibitor screening. Chemicon's Proteasome Activity Assay Kit provides a simple and convenient means for assaying proteasome activity that recognize the substrate LLVY (Meng et al. 1999). The assay is based on detection of the fluorophore 7-Amino-4-methylcoumarin (AMC) after cle…
