Revvity, 64RBM39TPEG, HTRF Human and Mouse Total RBM39 Detection Kit, 500 Assay Points

Overview
RNA-binding motif protein 39 (RBM39) is a pre-mRNA splicing factor involved in several biological processes, such as transcriptional regulation, alternative splicing, and protein translation. Upregulation of RBM39 indirectly participates in the growth and progression of tumors by regulating the transcription of many tumor-related genes, protein translation, and selective splicing, and its inhibition is lethal to several cancers including lung, breast, and colorectal cancer. For these reasons, the RBM39 protein has become an emerging target for cancer diagnosis and treatment.
How it works
Total-RBM39 assay principle
The HTRF Total-RBM39 assay quantifies the expression level of RBM39 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-RBM39 assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of RBM39 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-RBM39 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Total-RBM39 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total-RBM39 one-plate assay protocol
Detection of Total-RBM39 with HTRF reagents can be performed in a single plate used for culturing, stimulation, 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 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Unit Size-500 assay points