Western blot analysis was performed on nuclear enriched extracts (30 ug lysate) of PC-3 (Lane 1), MCF7 (Lane 2), MDA-MB-231 (Lane 3) and whole cell extracts (30 ug lysate) of SK-OV-3 (Lane 4) and PC-12 (Lane 5). The blots were probed with Anti-Estrogen Related Receptor Rabbit Polyclonal (Product # PA1-314, 1-3 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product #A27036, 0.4ug/ml, 1:2500 dilution). A 45 kDa band corresponding to Estrogen- Related Receptor alpha was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues M(1) S S Q V V G I E P L Y I K A E(16) of human ERR1.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1-3 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-314 detects human ERR1 from transfected cells and shows no cross-reactivity to ERR3.
PA1-314 has been successfully used in Western blot and immunohistochemistry procedures. By Western blot, this antibody detects an ~52 kDa protein representing ERR1 from COS-7 transfected cells. An unidentified band is seen at ~37 kDa.
The PA1-314 immunogen is a synthetic peptide corresponding to the residues M(1) S S Q V V G I E P L Y I K A E(16) of human ERR1. This sequence is completely conserved between canine, human, mouse and rat. This immunizing peptide (PEP-231) can be used for neutralization and control experiments.
Estrogen-related receptor alpha 1 (ERR alpha, ERR1) is an orphan member of the nuclear hormone receptor superfamily. All the ERR family members share an almost identical DNA-binding domain, which has 68% amino acid identity with that of estrogen receptor.
This protein binds to an ERR-alpha response element (ERRE) containing a single consensus half-site, 5'-tnaaggtca-3' can bind to the medium-chain acyl coenzyme a dehydrogenase (mcad) response element nrre-1 and may act as an important regulator of mcad promoter. Studies have shown that ERR1 functions a metabolic regulator and is often expressed in tissues that preferentially metabolize fatty acids.
The corresponding gene for the ERR1 is NR3B1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Identification and characterization of estrogen receptor-related receptor alpha and gamma in human glioma and astrocytoma cells.
PA1-314 was used in western blot to characterize estrogen receptor-related receptor alpha and gamma in glioma and astrocytoma cells
|Gandhari MK,Frazier CR,Hartenstein JS,Cloix JF,Bernier M,Wainer IW||Molecular and cellular endocrinology (315:314)||2010|