Revvity, 64CYCE1TPEH, HTRF Human Total Cyclin E1 Detection Kit, 10,000 Assay Points

Overview
E-type cyclins play an important role in the transition of quiescent cells into the cell cycle. Two E-type cyclins have been previously described, Cyclin E1 and Cyclin E2. Cyclin E1 is known to activate CDK2 inducing S-phase transition and DNA synthesis associated with mitosis. Overexpression of Cyclin E1 is directly implicated in many cancers.
This kit is compatible with the buffer from Total & phospho-CDK2 kits (‰ 64CDK2TPEG & ‰  64CDK2Y15PEG), so the same lysate can be used for analyses of the level of Total Cyclin E1, Total CDK2 and CDK2 phospho Tyr15.
How it works
Total Cyclin E1 assay principle
The Total Cyclin E1 assay quantifies the expression level of Cyclin E1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total Cyclin E1 assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In the presence of Cyclin E1 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein's expression under a no-wash assay format.

Total Cyclin E1 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a low volume detection plate before the addition of Total Cyclin E1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total Cyclin E1 one-plate assay protocol
Detection of Total Cyclin E1 with HTRF reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Unit Size-10,000 assay points