Revvity, 64SMARC2TPEG, HTRF Human Total SMARCA-2 Detection Kit, 500 Assay Points

Overview
The SMARCA subgroup of genes belongs to the SWI/SNF family, involved in chromatin remodeling and DNA damage repair. SWI/SNF complexes regulate the genes involved in essential processes such as DNA replication, damage repair, and cell proliferation.
SMARCA4/BRG1 and SMARCA2/BRM are two mutually exclusive catalytic ATPase subunits of the SWI/SNF complex.
SMARCA4 loss of function is frequently associated with cancer, and a synthetic lethal relationship between SMARCA4 and SMARCA2 is well established in SMARCA4-deficient cancers, where the inactivation of SMARCA2 leads to tumor cell death.
Among therapeutic strategies being investigated to fight cancers, targeted protein degradation is expected to overcome some limitations from conventional small molecules. So far, PROTAC molecules have been developed to induce the selective degradation of SMARCA proteins. However, SMARCA2 and SMARCA4 share about 75% identity at the protein level, and so compounds are likely to target both proteins. Therefore, the selectivity of compounds targeting either SMARCA2 or SMARCA4 must be carefully investigated. In such investigations, the HTRF total SMARCA4 kit is a valuable companion assay for the HTRF total SMARCA2 kit, enabling reliable compound profiling.
How it works
Total-SMARCA2 assay principle
The HTRF Total-SMARCA2 assay quantifies the expression level of SMARCA2 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-SMARCA2 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 SMARCA2 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-SMARCA2 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-SMARCA2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

Total-SMARCA2 one-plate assay protocol
Detection of Total-SMARCA2 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 2 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-500 assay points