Immunofluorescent analysis of Estrogen Receptor beta (ER-beta, green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 5% normal goat serum (Product # 31873) for 15 minutes at room temperature. Cells were probed with an ER-beta polyclonal antibody (Product # PA1-312) at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:200 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C(467) C S T E D S K N K E G S Q N L Q S Q(485) of mouse ER beta.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (IHC)||1-2 µg/ml|
|Western Blot (WB)||2 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 3 publications below|
PA1-312 detects estrogen receptor (ER) beta from rat, human, and mouse samples.
PA1-312 has been successfully used in Western blot, immunofluorescence, and immunohistochemistry procedures. By Western blot, this antibody detects an ~55 kDa protein representing ER beta from rat brain homogenate. Immunohistochemical staining of ER beta in rat cerebellum with PA1-312 results in staining of cells in the paraventricular nucleus and retrochiasmatic nucleus.
The PA1-312 immunogen is a synthetic peptide corresponding to residues C(467) C S T E D S K N K E G S Q N L Q S Q(485) of mouse ER beta.
Recombinant ER beta expressed in baculovirus system (Cat. # RP-311 and RP-312) can be purchased for control experiments in Western blot and gel shift assays.
The human ER-beta is a newly discovered estrogen receptor initially cloned and characterized from testis. The size and structure of ER-beta is very similar to ER-alpha with the ligand and DNA binding domains being highly conserved, while the amino terminus which serves as their transactivation domain has diverged significantly. Similar in function to ER-alpha ER-beta binds to estrogen with a high affinity and regulates estrogen specific gene activation through direct interaction with estrogen response elements (ERE's).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The Modifier of hemostasis (Mh) locus on chromosome 4 controls in vivo hemostasis of Gp6-/- mice.
PA1-312 was used in western blot to study the role of platelet glycoprotein VI and a new locus on chromosome 4 in platelet thrombus formation and homeostasis
|Cheli Y,Jensen D,Marchese P,Habart D,Wiltshire T,Cooke M,Fernandez JA,Ware J,Ruggeri ZM,Kunicki TJ||Blood (111:1266)||2008|
Genistein affects HER2 protein concentration, activation, and promoter regulation in BT-474 human breast cancer cells.
PA1-312 was used in western blot to study the effect of genistein on HER2 expression and function in BT-474 human breast cancer cells
|Sakla MS,Shenouda NS,Ansell PJ,Macdonald RS,Lubahn DB||Endocrine (32:69)||2007|
Phytoestrogens in common herbs regulate prostate cancer cell growth in vitro.
PA1-312 was used in western blot to investigate the effect of phytoestrogens on prostate tumor growth
|Shenouda NS,Zhou C,Browning JD,Ansell PJ,Sakla MS,Lubahn DB,Macdonald RS||Nutrition and cancer (49:200)||2004|