Active Motif,
Tools to analyze nuclear function,
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Gene Regulation

NR Sandwich ERα ELISA

ERα is a well-established marker of breast cancer hormone sensitivity, and the quantification of estrogen receptors in breast tumors has been routinely performed in clinical laboratories to aid in the selection between hormonal and chemotherapy and also to predict prognosis. Because of ERα's critical role in cell biology, it is important to measure the total amounts of ERα contained in different cell types and tissues.

 
Name Format Cat No. Price  
NR Sandwich ERα 1 x 96 rxns 49296 $695 Buy Now
5 x 96 rxns 49796 $2,850 Buy Now
 
 
 
Figure 1:Measuring cellular ERα levels.

Different amounts of nuclear extracts from MCF-7 and MDA-MB-231 cells were analyzed for cellular levels of ERα using the NR Sandwich ERα Kit.

Applications

The NR Sandwich ERα 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 sensitive, accurate quantification of the nuclear receptor of interest (NR).

Contents & Storage

One or five 96-well plate(s) coated with Capture Antibody, detecting antibody, HRP-conjugated secondary antibody, positive control cell extract and reaction buffers. Reagent storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months.