Product Pathways - Nuclear Receptor Signaling
PathScan® Total Estrogen Receptor α Sandwich ELISA Kit #80251
|Product Includes||Volume||Solution Color|
|ELISA Sample Diluent||25 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|STOP Solution #7002||11 ml||Colorless|
|TMB Substrate #7004||11 ml||Colorless|
|Estrogen Receptor α Mouse Detection mAb||1 ea||Green (Lyophilized)|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
|Estrogen Receptor α Rabbit mAb Coated Microwells||1 ea|
|Sealing Tape||2 sheets|
Specificity / Sensitivity
PathScan® Total Estrogen Receptor α Sandwich ELISA Kit detects endogenous levels of estrogen receptor α protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The PathScan® Total Estrogen Receptor α Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of estrogen receptor α protein. An Estrogen Receptor α Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-estrogen receptor α proteins are captured by the coated antibody. Following extensive washing, an Estrogen Receptor α Mouse Detection mAb is added to detect captured estrogen receptor α proteins. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for the developed color is proportional to the quantity of estrogen receptor α protein.
Antibodies in the kit are custom formulations specific to the kit.
ELISA - Western correlation
Figure 1. Estrogen receptor α protein is detected from multiple cell lines using the PathScan® Total Estrogen Receptor α Sandwich ELISA Kit #80251. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using Estrogen Receptor α (D6R2W) Rabbit mAb #13258 is shown in the bottom figure.
Estrogen receptor α (ERα), a member of the steroid receptor superfamily, contains highly conserved DNA binding and ligand binding domains (1). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ERα regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (2). Phosphorylation at multiple sites provides an important mechanism to regulate ERα activity (3-5). Ser104, 106, 118, and 167 are located in the amino-terminal transcription activation function domain AF-1, and phosphorylation of these serine residues plays an important role in regulating ERα activity. Ser118 may be the substrate of the transcription regulatory kinase CDK7 (5). Ser167 may be phosphorylated by p90RSK and Akt (4,6). According to the research literature, phosphorylation at Ser167 may confer tamoxifen resistance in breast cancer patients (4).
- Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
- Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
- Chen, D. et al. (1999) Mol Cell Biol 19, 1002-15.
- Campbell, R.A. et al. (2001) J Biol Chem 276, 9817-24.
- Chen, D. et al. (2000) Mol Cell 6, 127-37.
- Joel, P.B. et al. (1998) Mol Cell Biol 18, 1978-84.
- Burns, K.A. and Korach, K.S. (2012) Arch Toxicol 86, 1491-504.
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- 13258 Estrogen Receptor α (D6R2W) Rabbit mAb
- 7002 STOP Solution
- 7004 TMB Substrate
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 8644 Estrogen Receptor α (D8H8) Rabbit mAb
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9998 BSA
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
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