Revvity, 64NPMTPEH, HTRF Human and Mouse Total NPM1 Detection Kit, 10,000 assay points

Overview
NPM1 (Nucleophosmin 1) is a molecular chaperone involved in cell cycle regulation, ribosome biogenesis, cell proliferation, DNA damage repair, and apoptosis. Its phosphorylation on Thr199 by the CDK2/Cyclin E complex regulates mitosis, centrosome duplication, pre-mRNA processing, and G2/M cell cycle arrest through its interaction with p53/MDM2.
NPM1 functions both as an oncogene and a tumor suppressor. It is frequently overexpressed, altered, and rearranged, or sporadically deleted, in human cancers. NPM1 is also the most frequently mutated gene (NPM1c) in acute myeloid leukemia (AML).
How it works
Total NPM1 assay principle

The Total NPM1 assay quantifies the expression level of NPM1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total NPM1 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 NPM1 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 NPM1 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 HTRF Total NPM1 detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 
Total NPM1 one-plate assay protocol

Detection of Total NPM1 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 allows miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-NPM1
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Oncology
Unit Size-10,000 assay points