PA1-313

antibody from Invitrogen Antibodies

Targeting: ESR2 ER-beta, Erb, NR3A2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-313

Provider

Invitrogen Antibodies

Product name

Estrogen Receptor beta Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA1-313 has been successfully used in Western blot 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 prostate with PA1-313 results in nuclear staining, whereas nuclear staining may not be observed in human tissue.

Concentration

1 mg/mL

References

Submitted references

The absence of oestrogen receptor beta disturbs collagen I type deposition during Achilles tendon healing by regulating the IRF5-CCL3 axis.

Bian X, Liu T, Yang M, Gu C, He G, Zhou M, Tang H, Lu K, Lai F, Wang F, Yang Q, Gustafsson JÅ, Fan X, Tang K
Journal of cellular and molecular medicine 2020 Sep;24(17):9925-9935

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Insights into the Role of Estrogen Receptor β in Triple-Negative Breast Cancer.

Sellitto A, D'Agostino Y, Alexandrova E, Lamberti J, Pecoraro G, Memoli D, Rocco D, Coviello E, Giurato G, Nassa G, Tarallo R, Weisz A, Rizzo F
Cancers 2020 Jun 5;12(6)

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Interaction Proteomics Identifies ERbeta Association with Chromatin Repressive Complexes to Inhibit Cholesterol Biosynthesis and Exert An Oncosuppressive Role in Triple-negative Breast Cancer.

Alexandrova E, Giurato G, Saggese P, Pecoraro G, Lamberti J, Ravo M, Rizzo F, Rocco D, Tarallo R, Nyman TA, Collina F, Cantile M, Di Bonito M, Botti G, Nassa G, Weisz A
Molecular & cellular proteomics : MCP 2020 Feb;19(2):245-260

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Absence of estrogen receptor beta leads to abnormal adipogenesis during early tendon healing by an up-regulation of PPARγ signalling.

Bian X, Liu T, Zhou M, He G, Ma Y, Shi Y, Wang Y, Tang H, Kang X, Yang M, Gustafsson JÅ, Fan X, Tang K
Journal of cellular and molecular medicine 2019 Nov;23(11):7406-7416

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Effect of estradiol on fibroblasts from postmenopausal idiopathic carpal tunnel syndrome patients.

Yamanaka Y, Menuki K, Tajima T, Okada Y, Kosugi K, Zenke Y, Sakai A
Journal of cellular physiology 2018 Nov;233(11):8723-8730

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Targeting Estrogen Receptor Beta in a Phase 2 Study of High-Dose Estradiol in Metastatic Triple-Negative Breast Cancer: A Wisconsin Oncology Network Study.

Wisinski KB, Xu W, Tevaarwerk AJ, Saha S, Kim K, Traynor A, Dietrich L, Hegeman R, Patel D, Blank J, Harter J, Burkard ME
Clinical breast cancer 2016 Aug;16(4):256-61

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Prognostic significance of full-length estrogen receptor beta expression in stage I-III triple negative breast cancer.

Shanle EK, Onitilo AA, Huang W, Kim K, Zang C, Engel JM, Xu W, Wisinski KB
American journal of translational research 2015;7(7):1246-59

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Histone H3 Lysine 27 demethylases Jmjd3 and Utx are required for T-cell differentiation.

Manna S, Kim JK, Baugé C, Cam M, Zhao Y, Shetty J, Vacchio MS, Castro E, Tran B, Tessarollo L, Bosselut R
Nature communications 2015 Sep 2;6:8152

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Characterization of estrogen response element binding proteins as biomarkers of breast cancer behavior.

Kruer TL, Cummins TD, Powell DW, Wittliff JL
Clinical biochemistry 2013 Nov;46(16-17):1739-46

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Estrogen receptor beta in cancer: an attractive target for therapy.

Gallo D, De Stefano I, Grazia Prisco M, Scambia G, Ferrandina G
Current pharmaceutical design 2012;18(19):2734-57

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Amounts of an estrogen receptor β isoform increased in the theca of preovulatory follicles of sheep.

Cárdenas H, Pope WF
Animal reproduction science 2012 Apr;131(3-4):143-52

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Morphologic transformation of human breast epithelial cells MCF-10A: dependence on an oxidative microenvironment and estrogen/epidermal growth factor receptors.

Yusuf R, Frenkel K
Cancer cell international 2010 Sep 1;10:30

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Estrogen increases survival in an orthotopic model of glioblastoma.

Barone TA, Gorski JW, Greenberg SJ, Plunkett RJ
Journal of neuro-oncology 2009 Oct;95(1):37-48

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Prepubertal physical activity up-regulates estrogen receptor beta, BRCA1 and p53 mRNA expression in the rat mammary gland.

Wang M, Yu B, Westerlind K, Strange R, Khan G, Patil D, Boeneman K, Hilakivi-Clarke L
Breast cancer research and treatment 2009 May;115(1):213-20

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Estrogen receptor beta displays cell cycle-dependent expression and regulates the G1 phase through a non-genomic mechanism in prostate carcinoma cells.

Hurtado A, Pinós T, Barbosa-Desongles A, López-Avilés S, Barquinero J, Petriz J, Santamaria-Martínez A, Morote J, de Torres I, Bellmunt J, Reventós J, Munell F
Cellular oncology : the official journal of the International Society for Cellular Oncology 2008;30(4):349-65

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Changes in biomarkers of estrogen receptor and growth factor signaling pathways in MCF-7 tumors after short- and long-term treatment with soy and flaxseed.

Power KA, Chen JM, Saarinen NM, Thompson LU
The Journal of steroid biochemistry and molecular biology 2008 Nov;112(1-3):13-9

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Differing transcriptional responses to pulsed or continuous estradiol exposure in human umbilical vein endothelial cells.

Li J, Wang H, Johnson SM, Horner-Glister E, Thompson J, White IN, Al-Azzawi F
The Journal of steroid biochemistry and molecular biology 2008 Jul;111(1-2):41-9

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Changes in transcription profile and cytoskeleton morphology in pelvic ligament fibroblasts in response to stretch: the effects of estradiol and levormeloxifene.

Ewies AA, Elshafie M, Li J, Stanley A, Thompson J, Styles J, White I, Al-Azzawi F
Molecular human reproduction 2008 Feb;14(2):127-35

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Estrogen receptors in the uterus and ovarian follicles of gilts treated with dihydrotestosterone.

Cárdenas H, Pope WF
Domestic animal endocrinology 2005 Oct;29(3):523-33

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Nuclear estrogen receptor beta in lung cancer: expression and survival differences by sex.

Schwartz AG, Prysak GM, Murphy V, Lonardo F, Pass H, Schwartz J, Brooks S
Clinical cancer research : an official journal of the American Association for Cancer Research 2005 Oct 15;11(20):7280-7

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Binding of estrogen receptor beta to estrogen response element in situ is independent of estradiol and impaired by its amino terminus.

Huang J, Li X, Maguire CA, Hilf R, Bambara RA, Muyan M
Molecular endocrinology (Baltimore, Md.) 2005 Nov;19(11):2696-712

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Single-chain estrogen receptors (ERs) reveal that the ERalpha/beta heterodimer emulates functions of the ERalpha dimer in genomic estrogen signaling pathways.

Li X, Huang J, Yi P, Bambara RA, Hilf R, Muyan M
Molecular and cellular biology 2004 Sep;24(17):7681-94

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Gonadal hormones and frontocortical expression of vascular endothelial growth factor in male stroke-prone, spontaneously hypertensive rats, a model for attention-deficit/hyperactivity disorder.

Jesmin S, Togashi H, Sakuma I, Mowa CN, Ueno K, Yamaguchi T, Yoshioka M, Kitabatake A
Endocrinology 2004 Sep;145(9):4330-43

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Prepubertal estradiol and genistein exposures up-regulate BRCA1 mRNA and reduce mammary tumorigenesis.

Cabanes A, Wang M, Olivo S, DeAssis S, Gustafsson JA, Khan G, Hilakivi-Clarke L
Carcinogenesis 2004 May;25(5):741-8

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Oestrogen receptor beta: what it means for patients with breast cancer.

Speirs V, Carder PJ, Lane S, Dodwell D, Lansdown MR, Hanby AM
The Lancet. Oncology 2004 Mar;5(3):174-81

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Estrogen receptor beta localization in the lizard (Podarcis s. sicula) testis.

Chieffi P, Varriale B
Zygote (Cambridge, England) 2004 Feb;12(1):39-42

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Estrogen regulates adrenal angiotensin AT1 receptors by modulating AT1 receptor translation.

Wu Z, Maric C, Roesch DM, Zheng W, Verbalis JG, Sandberg K
Endocrinology 2003 Jul;144(7):3251-61

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17Beta-estradiol up-regulates vascular endothelial growth factor receptor-2 expression in human myometrial microvascular endothelial cells: role of estrogen receptor-alpha and -beta.

Gargett CE, Zaitseva M, Bucak K, Chu S, Fuller PJ, Rogers PA
The Journal of clinical endocrinology and metabolism 2002 Sep;87(9):4341-9

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17-Beta-estradiol regulates expression of genes that function in macrophage activation and cholesterol homeostasis.

Kramer PR, Wray S
The Journal of steroid biochemistry and molecular biology 2002 Jul;81(3):203-16

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17-Beta-estradiol regulates expression of genes that function in macrophage activation and cholesterol homeostasis.

Kramer PR, Wray S
The Journal of steroid biochemistry and molecular biology 2002 Jul;81(3):203-16

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Differences in the abilities of estrogen receptors to integrate activation functions are critical for subtype-specific transcriptional responses.

Yi P, Bhagat S, Hilf R, Bambara RA, Muyan M
Molecular endocrinology (Baltimore, Md.) 2002 Aug;16(8):1810-27

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Expression of estrogen receptors alpha and beta in the baboon fetal ovary.

Pepe GJ, Billiar RB, Leavitt MG, Zachos NC, Gustafsson JA, Albrecht ED
Biology of reproduction 2002 Apr;66(4):1054-60

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Loss of estrogen receptor beta expression in malignant human prostate cells in primary cultures and in prostate cancer tissues.

Pasquali D, Rossi V, Esposito D, Abbondanza C, Puca GA, Bellastella A, Sinisi AA
The Journal of clinical endocrinology and metabolism 2001 May;86(5):2051-5

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Different regulation of oestrogen receptors alpha and beta in the human cervix at term pregnancy.

Wang H, Stjernholm Y, Ekman G, Eriksson H, Sahlin L
Molecular human reproduction 2001 Mar;7(3):293-300

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Estrogen receptors in the normal adult and developing human inner ear and in Turner's syndrome.

Stenberg AE, Wang H, Fish J 3rd, Schrott-Fischer A, Sahlin L, Hultcrantz M
Hearing research 2001 Jul;157(1-2):87-92

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Estradiol acts as a germ cell survival factor in the human testis in vitro.

Pentikäinen V, Erkkilä K, Suomalainen L, Parvinen M, Dunkel L
The Journal of clinical endocrinology and metabolism 2000 May;85(5):2057-67

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Estrogen receptor beta is coexpressed with ERalpha and PR and associated with nodal status, grade, and proliferation rate in breast cancer.

Järvinen TA, Pelto-Huikko M, Holli K, Isola J
The American journal of pathology 2000 Jan;156(1):29-35

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Identification and developmental expression of the estrogen receptor alpha and beta in the baboon fetal adrenal gland.

Albrecht ED, Babischkin JS, Davies WA, Leavitt MG, Pepe GJ
Endocrinology 1999 Dec;140(12):5953-61

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of T-47 D (Lane 1), MCF7 (Lane 2), MDA-MB-231 (Lane 3), SK-BR-3 (Lane 4), tissue extracts of Rat Ovary (Lane 5), Rat Testis (Lane 6), Rat Colon (Lane 7), Rat Brain (Lane 8) and Mouse Brain (Lane 9). The blot was probed with Anti-Estrogen Receptor beta Polyclonal Antibody (Product # PA1-313, 1µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 67 kDa band corresponding to Estrogen Receptor alpha and 55 kDa band corresponding to Estrogen Receptor beta was observed across the cell lines and tissues tested.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ER beta was performed by loading the indicated amounts of recombinant full-length human ER alpha (Product # A15674) and ER beta (Product # A15664) proteins, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temp. ER beta was detected at ~54 kD (left panel) using an ER beta polyclonal antibody (Product # PA1-313) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). The blot was then stripped with Restore Plus Western blot stripping buffer (Product # 46430) for at least 15 min at room temp, re-blocked with 5% milk in TBST for at least 1 hour at room temp, and re-probed with an ER alpha polyclonal antibody (Product # PA1-308, right panel) at a dilution of 1:2000 overnight at 4C, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). The presence of faint ER-beta bands on the blot probed with PA1-308 is due to incomplete stripping of PA1-313.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of T-47D (Lane 1), MCF7 (Lane 2), MDA-MB-231 (Lane 3), and SK-BR-3 (Lane 4). The blot was probed with Anti-Estrogen Receptor beta Polyclonal Antibody (Product # PA1-313, 1 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4,000 dilution). A 55 kDa band corresponding to Estrogen Receptor beta was observed across the cell lines and tissues tested along with an uncharacterized band (*) at ~70 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Estrogen Receptor beta was performed using 70% confluent log phase T-47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Estrogen Receptor beta Polyclonal Antibody (Product # PA1-313) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunolocalization of ER beta in rat prostate using Product # PA1-313.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

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Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

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Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

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Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

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Experimental details

Figure 7 Analysis of MCF-10A Cells for ERalpha, ERbeta, and EGFR . Nuclear proteins extracted from MCF-10A cells that were exposed to differing culture conditions were analyzed by Western blotting for the presence of ERalpha, ERbeta, and EGFR. Numbers in parentheses indicate the number of passages in that particular type of medium. Proteins (42 mug/lane) were electrophoresed onto two 12% Tris-HCl gels (lanes 1-8 and 9-16 respectively) and transferred onto nitrocellulose membranes. Membranes were probed with antibodies directed against ERalpha (a and b), stripped and re-probed with antibodies directed against ERbeta (c and d), and stripped a second time and re-probed with antibodies directed against EGFR (e and f). Results shown are those from one experiment.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of ERalpha and ERbeta in the SSCT derived from idiopathic patients (magnification x400). (a) Negative control. (b) ERalpha expression (indicated by an arrow). (c) ERbeta expression (indicated by an arrow). (d) ERalpha expression was higher than ERbeta expression, but this difference was not significant. All results are expressed as means +- standard error

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 RNA-seq analysis of the gene expression profile of ERbeta -/- and WT control tendon scars revealed IRF5 down-regulation in the absence of ERbeta. A, Heatmap depicting the expression levels of genes in WT and ERbeta -/- tendons after 1 wk of repair. B, Volcano map of differentially expressed genes between WT and ERbeta -/- tendons after 1 wk of repair: 68 genes were up-regulated, and 51 genes were down-regulated. C, Heatmap of differentially expressed transcription factors. D, Immunoblots of IRF5 in WT and ERbeta -/- tendons after 1 wk of repair. E, Bioinformatic prediction of binding sites of ERbeta and IRF5 using JASPAR (chr6:29528943-29528949, AGGGCAG, length = 7). F, The immunofluorescence analysis of ERbeta and IRF5 revealed co-expression in macrophages. Scale bars: 50 mum

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1 ERbeta -/- tendon scars have an inferior tendon repair process. A-D, Surgery process of mouse Achilles tendon. E, F Low-magnification HE staining indicates a very different scar organization with clear adipocyte accumulation in ERbeta -/- mice. (G,H) Cell density in the healing region was significantly lower in ERbeta -/- mice compared with WT controls. (I,J) Number of CD34-labelled blood vessels was significantly higher in ERbeta -/- mice compared with WT controls. K, Evaluation of tendon healing using an established histological scoring system revealed that ERbeta -/- mice had a significantly lower histological score compared with WT controls. Data are represented as mean +- SEM (n = 5), * P < .05, ** P < .01. Scale Bars: 100mum

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 2 ERbeta -/- tendon scars have reduced cell proliferation, whilst increased cell apoptosis. A-C, Immunohistochemistry of ERbeta revealed a little expression in normal mouse Achilles tendon (A), more expression in early healing of WT mouse Achilles tendon (B) and no expression in ER-beta -/- mouse injured Achilles tendon. ERbeta -/- mouse Achilles tendon (C). (D,G,F) TUNEL staining and statistical analysis showed more apoptotic cells in ERbeta -/- tendon scars. (E,H,I) Immunohistochemistry of Ki67 and statistical analysis revealed decreased cell proliferation in ERbeta -/- tendon scars. Data are represented as mean +- SEM (n = 5), *** P < .001. Scale Bars: 100mum