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Cereblon (CRBN) is a ubiquitously expressed protein which functions as a substrate recognition subunit within the CRL4-CRBN E3 ubiquitin ligase complex and targets specific proteins for proteolysis through the ubiquitin proteasome system (UPS). Cereblon plays a key role in normal limb outgrowth, normal cell proliferation and metabolism, and in memory and learning. Cereblon is also the primary direct target of the immunomodulatory drugs Thalidomide, Pomalidomide, and Lenalidomide, which are FDA approved for treating multiple myeloma. Like other E3 ligases such as VHL, Cereblon E3 ligase property is commonly used in targeted protein degradation strategy, eg PROTAC.
AlphaLISA technology allows the detection of molecules of interest in a no- wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
This product is also available using HTRF Technology. To learn more, visit HTRF Cereblon Binding Kit webpage.
PerkinElmer has made available a range of documents with educational content and reagents dedicated to PROTAC to help you in your research. Visit our dedicated webpage to learn more.
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Therapeutic Area||Central Nervous System, Oncology|
|Unit Size||5,000 Assay Points|
Alpha (Amplified Luminescent Proximity Homogeneous Assay) technology is a bead-based, no-wash alternative to traditional ELISAs. Instead of detection with an HRP substrate, the Alpha assay signal is generated by the excitation of an Eu+-coated bead that has been conjugated to the detection antibody.
Alpha technology offers a simple, straight forward workflow. No wash needed!
Download this application note to see how no-wash AlphaLISA® technology provides a more-convenient alternative to ELISA for quantitation of biomarkers in complex sample types, including tissue, serum, and plasma.
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.