MA1-310

antibody from Invitrogen Antibodies

Targeting: ESR1 ER-alpha, Era, ESR, NR3A1

Provider product page for MA1-310

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [48]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Other assay [12]

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Product number: MA1-310 - Provider product page
Provider: Invitrogen Antibodies
Product name: Estrogen Receptor alpha Monoclonal Antibody (33)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA1-310 detects both the steroid occupied and unoccupied forms of the estrogen receptor (ER) alpha from human samples. MA1-310 detects ER alpha and does not cross react with ER beta. MA1-310 has been successfully used in Western blot, immunohistochemistry, immunoprecipitation, gel shift, flow cytometry, and immunocytochemical experiments. By Western blot, this antibody detects an ~66 kDa protein representing ER alpha from MCF-7 cell extract. MA1-310 can be used with blocking peptide PEP-013.
Reactivity: Human, Mouse, Rat, Bovine
Host: Mouse
Isotype: IgG
Antibody clone number: 33
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

ESR1 protein structure - MA1-310 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Estrogen Receptor Alpha was performed by loading 25 µg of the indicated whole cell lysates and 10 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. Estrogen Receptor Alpha was detected at 66 kD using a Estrogen Receptor Alpha mouse monoclonal antibody (Product # MA1-310) at a concentration of 4 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Superclonal goat anti-mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Estrogen Receptor (ER) Alpha 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 Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% Milk in TBST for at least 1 hour at room temperature. ER Alpha was detected at 66 kD (left panel) using an ER Alpha mouse monoclonal antibody (Product # MA1-310) at a concentration of 4 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Superclonal goat anti-mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076). Directly alongside the above blot, a second blot containing the same recombinant ER alpha and ER beta protein samples was probed with an ER-beta specific polyclonal antibody (Product # PA1-313, right panel) at a dilution of 1:2000 overnight at 4C, followed by an HRP-conjugated Superclonal goat anti-rabbit IgG secondary antibody (Product # A27034) at a dilution of 1:5000. Chemiluminescent detection was also performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 VAMP7 enhances estrogen receptor transcriptional activity. ( a ) Luciferase assays following transfection with VAMP7 or ESR1 or VAMP7 and ESR1 in HeLa cells incubated in absence (EtOH) or presence of 17 beta-estradiol (10 -8 M) for 24 h. n = 3 independent experiments for each condition. Data are presented as means +- s.e.m. One-way analysis of variance (ANOVA) with post hoc Bonferroni test was used for statistical analyses. * P < 0.05; ** P < 0.01; *** P < 0.001. ( b ) Western Blot analysis of ESR1 and LMNA (lamin A/C) in nuclear protein extracts of HeLa cells co-transfected with ESR1 or ESR1 and VAMP7 in absence (EtOH) or presence of 17 beta-estradiol (10 -8 M) for 24 h. ( c ) Reciprocal co-immunoprecipitation of ESR1 and VAMP7 following their co-transfection in HeLa cells. ( d ) qRT-PCR analysis of key genes of ESR1 signaling in testis from WT ( n = 3) and V7BAC mice (line 7; n = 3). Data are expressed as mean +- s.e.m. Mean differences between WT and V7BAC were determined by unpaired, two-tailed Student's t -test. * P < 0.05; ** P < 0.01; *** P < 0.001. ( e ) ATF3 immunostaining of testis and external genitalia at fetal and adult stages of WT and V7BAC mice. For fetal tissues: scale bar, 250 um. For adult urethra and testis: scale bar, 100 um. ( f ) qRT-PCR of VAMP 7 and ATF3 gene expression after incubation with non-targeting (scramble) or VAMP7 siRNA in NT2/D1. n = 3 independent experiments for each condition. Mean differences were determined by unpaired, t

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 1 Estrogen receptor (ER) staining intensities of endothelial cells. Boxplot graphic of ERalpha and ERbeta. Group 1, OVX; Group 2, OVX+RL; Group 3, OVX+AV; Group 4, OVX+RL+AV; Data are presented as median. Kruskal-Wallis test was performed to determine the differences between groups.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 FZS and E2 show discrete effects on male and female OPCs. Western blot results showing the expression of ( A ) MAG, ( B ) Nup133 and ( C ) ERalpha in OPCs after treatment with 100 nM Adiol (AD), 100 nM Allopregnanolone (AL) and 100 nM 16alpha-OH-DHEA under normoxia and hyperoxia. All data are representative of at least three independent experiments. Bars and error represent mean +- SEM of replicate measurements. Significant differences from normoxia control are indicated by * p < 0.05 (Student's t -test).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 mRNA and protein expression of AKR1C1 ( A , E ), P450 ( B , F ), 3beta-HSD ( C , G ) and 17beta-HSD ( D , H ) in uterine tissues (endometrium and myometrium) on 4th and 13th day of estrous cycle, in pregnancy and anestrus phase. Data were normalized against GAPDH for mRNA expression and against beta-actin (ACTB) for proteins expression. Each bar represents one experimental group with SEM. Statistical differences were analyzed by two-way analysis (ANOVA) of variance followed by the Bonferroni post hoc test using GraphPad PRISM (Version 8.3.0). The lowest statistical significance was P < 0.05. Asterisks indicate statistical differences between endometrium and myometrium (* P < 0.1; ** P < 0.01; *** P < 0.001; **** P < 0.0001). Different letters indicate statistical differences ( P < 0.05) between the experimental groups throughout endometrium (A, B) and myometrium (a-b) respectively.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Nuclear aromatase expression overlaps with nuclear ERalpha signal. Double immunostaining with ERalpha (green) and Aro (red) in glia culture ( a - l ) shows that nuclear Aro signals colocalize with nuclear ERalpha ( a , e , i ; white arrowheads). Both Aro and ERalpha are strongly represented in the nucleus ( b , c , f , g , j,k ; yellow arrowheads) and also in the cytoplasm ( b , c , f , g , j , k ; blue arrowheads). ERalpha positive dots are also detectable in the processes ( b , f,j ; orange arrowheads). Scale bar: 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Silencing SHP2 expression induces acini-like structure formation. A) Light pictures of parental, control and SHP2-silenced MDA-MB231 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. B) Light pictures of parental, control and SHP2-silenced MDA-MB468 cells cultured in 3D LRBM matrigel that allows acini-like structure formation. Pictures of acini-like structures formed by the MCF-10A cells cultured under identical conditions is shown in both A and B for comparison. C) Effect of SHP2 silencing on expression of basal and luminal markers and estrogen receptor alpha (ERalpha). Data presented is representative of at least three independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Effect of ER siRNA on human ASM proliferation and ER Protein Expression MTT assay was used to study the effect of ERβ agonist WAY and ER&alpha agonist PPT on PDGF-induced proliferation in Asthmatic and Non-Asthmatic human ASM cells transfected with ERβ siRNA (Figure 5A) and ERα siRNA (Figure 5B) respectively. There was significant mitogenic effect observed in ERβ siRNA transfected cells compared to negative siRNA in both asthmatic and non-asthmatic ASM cells. ERβ siRNA transfected human ASM cells treated with WAY and WAY+PDGF produced a significant increase in proliferation as compared to negative siRNA treated group. PPT+PDGF treatment produced an increased proliferation in ERα siRNA transfected human ASM cells. Western blot analysis was performed to determine the transfection efficacy of ERβ (Figure 5C) and ERα (Figure 5D) receptor siRNA in human ASM cells. Values are means SEM from n of 5 samples from asthmatics and n of 6 samples from non-asthmatics for MTT assay. Values are means SEM from n of 3 different patient samples of asthmatics and non-asthmatics for western analysis. ***p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Histopathological and molecular characteristics of bone metastases in breast cancer, following intra-femoral inoculation of syngeneic MRMT-1 tumor cells. ( A ) Sham-operated female rats display a healthy femoral bone marrow (BM) and cortical bone (Cb) while ( B ) tumor-bearing rats exhibit intense cancer cell invasion into the bone marrow, 18 days following cancer cell implantation. ( C ) Extensive Ki-67-positive staining (brown-stained cancer cell nuclei) indicates a highly proliferative tumor, also expressing ERalpha (estrogen alpha receptor) ( D ) but not the HER-2 receptor ( E ). An anonymous human HER-2 breast cancer sample is used here as a positive control for HER-2 staining ( F ). Scale bars represent 100 um in ( A , B ), 200 um in ( C ) and 50 um in ( D-F ).