Figure 1:Measuring cellular ER? levels.
Different amounts of nuclear extracts from MCF-7 and MDA-MB-231 cells were analyzed for cellular levels of ER? using the NR Sandwich ER? Kit.
The NR Sandwich ER? has many applications including:
- Study of ER transcriptional activity regulation
- Protein structure/function studies of ER and its mutated variants in areas such as osteoporosis, arteriosclerosis and breast cancer
Nuclear receptors (NR) belong to a superfamily of structurally related ligand-inducible transcription factors whose members include receptors for estrogen (ER), glucocorticoid (GR), androgen (AR), thyroid hormone (TR), progesterone (PR) and vitamin D (VDR). Inappropriate signaling by these receptors is associated with numerous disease states, including cancer, asthma, infertility, osteoporosis and arthritis.
Why use the Nuclear Receptor ELISAs?
- Quantitative results in just hours
- ELISA format eliminates the need to run, blot and develop gels
- NR Sandwich measures Total Nuclear Receptor
- Analyze multiple samples in low volume, high throughput
- Ability to assay both cellular extracts and recombinant proteins
The NR Sandwich ELISA method
In order to fully examine the activation of nuclear receptors, it is important to be able to quantify the total levels of a given nuclear receptor within a sample. Traditionally, total nuclear receptor protein levels have been monitored using Western blot, EMSA and reporter assays, which can be time-consuming and difficult to quantify. Active Motif's NR Sandwich Kits provide a simple, rapid and quantitative alternative to traditional low-throughput, labor-intensive assays. The NR Sandwich ELISAs use two antibodies that recognize different epitopes on the protein of interest. The Capture Antibody, which is supplied immobilized in the wells of an ELISA plate, binds the nuclear receptor protein when the sample is added. The Detecting Antibody then binds to the captured nuclear receptor protein. Subsequent incubation with a HRP-conjugated secondary antibody provides an easily quantified readout of total nuclear receptor protein levels (Figure 1).
Figure 1: NR Sandwich ELISA schematic.
Capture and Detecting Antibodies are used for