ER alpha (5G7) Monoclonal Antibody

$Western blot analysis was performed on 100 fmol ER alpha (ERa) and ER beta (ERb1) recombinant protein with ER alpha (5G7) Monoclonal Antibody (bsm-53007M) at a concentration of 7 \u03bcg\/ ml. Figure shows that the antibody is specific for ER alpha isoform, whereas the ER beta isoform is not recognized.$

Western blot analysis was performed on 100 fmol ER alpha (ERa) and ER beta (ERb1) recombinant protein with ER alpha (5G7) Monoclonal Antibody (bsm-53007M) at a concentration of 7 \u03bcg\/ ml. Figure shows that the antibody is specific for ER alpha isoform, whereas the ER beta isoform is not recognized.

$COS-7 cells transiently over-expressing human ER\u03b1 (left) or ER\u00df1 (right) were labeled with the ER alpha (5G7) Monoclonal Antibody (bsm-53007M), used at a concentration of 15 \u03bcg\/ml, followed by a biotinylated secondary antibody and peroxidase-labeled avidin. Figure shows the specificity of the antibody for the ER alpha isoform.$

COS-7 cells transiently over-expressing human ER\u03b1 (left) or ER\u00df1 (right) were labeled with the ER alpha (5G7) Monoclonal Antibody (bsm-53007M), used at a concentration of 15 \u03bcg\/ml, followed by a biotinylated secondary antibody and peroxidase-labeled avidin. Figure shows the specificity of the antibody for the ER alpha isoform.

$ChIP assays was performed using MCF7 cells treated with the ER agonist estradiol for 3 hours prior to harvesting, ER alpha (5G7) Monoclonal Antibody (bsm-53007M), and optimized primer sets for qPCR. Sheared chromatin from 3 million cells and 5 \u03bcg of antibody were used per ChIP experiment. Recovery (%: ChIP\/input) and occupancy (x fold: +ve\/-ve) are shown. QPCR was performed with primers for the GREB1 promoter and for exon 2 of the myoglobin gene used as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +\/- control target). These results demonstrate the occupancy of the GREB1 promoter by ERalpha.$

ChIP assays was performed using MCF7 cells treated with the ER agonist estradiol for 3 hours prior to harvesting, ER alpha (5G7) Monoclonal Antibody (bsm-53007M), and optimized primer sets for qPCR. Sheared chromatin from 3 million cells and 5 \u03bcg of antibody were used per ChIP experiment. Recovery (%: ChIP\/input) and occupancy (x fold: +ve\/-ve) are shown. QPCR was performed with primers for the GREB1 promoter and for exon 2 of the myoglobin gene used as a negative control. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +\/- control target). These results demonstrate the occupancy of the GREB1 promoter by ERalpha.

Applications

WB
IHC-P
IHC
dot-blot

Reactivity

Human
Monkey

Overview
Catalog #	bsm-53007M
Product Name	ER alpha (5G7) Monoclonal Antibody
Applications	WB IHC-P IHC DOT-BLOT
Reactivity	Human, Monkey
Specifications
Conjugation	Unconjugated
Host	Mouse
Source	Monoclonal antibody raised in mouse against the NH2 terminus of the human ER alpha (estrogen receptor alpha), using a KLH-conjugated synthetic peptide (Q19-K32).
Clonality	Monoclonal
Clone #	5G7
Isotype	IgG3
Concentration	2ug/ul
Purification	Monoclonal antibody in PBS containing 0.05% azide; purified by ammonium sulphate precipitation followed by dialysis
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Store at -20C for 12 months.
Target
Gene ID	2099
Swiss Prot	P03372
Synonyms	ESR, ESR1, ESRA, NR3A1
Background	Nuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Ligand-dependent nuclear transactivation involves either direct homodimer binding to a palindromic estrogen response element (ERE) sequence or association with other DNA-binding transcription factors, such as AP-1/c-Jun, c-Fos, ATF-2, Sp1 and Sp3, to mediate ERE-independent signaling. Ligand binding induces a conformational change allowing subsequent or combinatorial association with multiprotein coactivator complexes through LXXLL motifs of their respective components. Mutual transrepression occurs between the estrogen receptor (ER) and NF-kappa-B in a cell-type specific manner. Decreases NF-kappa-B DNA-binding activity and inhibits NF-kappa-B-mediated transcription from the IL6 promoter and displace RELA/p65 and associated coregulators from the promoter. Recruited to the NF-kappa-B response element of the CCL2 and IL8 promoters and can displace CREBBP. Present with NF-kappa-B components RELA/p65 and NFKB1/p50 on ERE sequences. Can also act synergistically with NF-kappa-B to activate transcription involving respective recruitment adjacent response elements; the function involves CREBBP. Can activate the transcriptional activity of TFF1. Also mediates membrane-initiated estrogen signaling involving various kinase cascades. Isoform 3 is involved in activation of NOS3 and endothelial nitric oxide production. Isoforms lacking one or several functional domains are thought to modulate transcriptional activity by competitive ligand or DNA binding and/or heterodimerization with the full length receptor. Essential for MTA1-mediated transcriptional regulation of BRCA1 and BCAS3. Isoform 3 can bind to ERE and inhibit isoform 1.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC
dot-blot