Western blot analysis of human placenta extract probed with Mouse anti Human Cytochrome P450 Aromatase (MCA2077S) followed by F(ab′)2 Rabbit anti Mouse IgG:HRP (STAR13B)

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 use for the detection of aromatase in KGN cells by western blotting
Image caption:
Effect of H89 on aromatase and BRCA1 expression in KGN cells. A, The human ovarian granulosa cell line KGN were treated with control (DMSO), 20 mM H89, 50 mM PD98059, 200 nM Wortmannin, 100 nM Rapamycin 100 nM, and cells were harvested 24 hours after treatment. Immunoblotting was performed using specific antibodies against aromatase, BRCA1 and a-tubulin; B, Aromatase and BRCA1 mRNA levels were measured by real-time RT-PCR. Gapdh was used for normalizing the real time PCR results. The data are expressed as a fold (Aromatase mRNA), or a percentage (BRCA1 mRNA) relative to the DMSO-treated control.

From: Ghosh S, Lu Y, Hu Y. A Role of CREB in BRCA1 Constitutive Promoter Activity and Aromatase Basal Expression. Int J Biomed Sci. 2008 Dec 15;4(4):260-265.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detaction of aromatase in human tissues by Immunohistochemistry on paraffin sections
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P450 arom immunoreactivity in testicular region adjacent to seminoma and in controls A: Intense aromatase immunostaining in IGCN cells. B: Placental-like alkaline phosphatase staining of IGCN basal cells C: Strong aromatase immunoreactivity of interstitial Leydig cells in normal testis (Lc) D: Intense immunostaining of luteal cells (Luc) in ovarian tissue Scale bars: A-B, 8 μm; C,12.5 μm; D, 5 μm.

From: Rago V, Romeo F, Aquila S, Montanaro D, Andò S, Carpino A. Cytochrome P450 aromatase expression in human seminoma. Reprod Biol Endocrinol. 2005 Dec 22;3:72.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detaction of aromatase in human tissues by Immunohistochemistry on paraffin sections
Image caption:
Morphology and P450 arom immunoreactivity of tumoral region in human testis with seminoma. A-B: Haematoxylin-eosin staining. C-D: Strong P450 arom immunoreactivity in cytoplasm of neoplastic cells (Nc) and unstained lymphocytes (L). Insert: absorption control. Scale bars: A, 20 μm; B-C, 12.5 μm; D, 5 μm.

From: Rago V, Romeo F, Aquila S, Montanaro D, Andò S, Carpino A. Cytochrome P450 aromatase expression in human seminoma. Reprod Biol Endocrinol. 2005 Dec 22;3:72.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detection of aromatase in human synovial tissue by immunohistochemistry on cryosections
Image caption:
Aromatase expression in synovial tissue and endogenous steroid hormone release from superfused synovium. (a) Immunohistochemistry of aromatase in one OA and one RA patient. Using the respective control antibody revealed no staining of positive cells (not shown). Magnification: 400×. (b) Density of aromatase-positive cells in OA (open bars, n = 20) and RA patients (hatched bars, n = 16). (c) Spontaneously released E2, E3, and free testosterone from standardized superfused pieces of synovial tissue of OA (open bars, n = 20) and RA patients (hatched bars, n = 18). (b,c) Values are given as box blots with the 5th, 25th, 50th (median), 75th, and 95th percentile when applicable. OA, osteoarthritis; RA, rheumatoid arthritis. Other abbreviations are as given in the legend to Fig. 1.

From: Schmidt M, Weidler C, Naumann H, Anders S, Schölmerich J, Straub RH. Androgen conversion in osteoarthritis and rheumatoid arthritis synoviocytes--androstenedione and testosterone inhibit estrogen formation and favor production of more potent 5alpha-reduced androgens. Arthritis Res Ther. 2005;7(5):R938-48.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detection of aromatase in porcine spermatozoa by immunofluorescence
Image caption:
Immunofluorescence labelling of androgen and estrogen receptors in pig spermatozoa: A) AR red fluorescence in sperm proximal mid-piece. B) P450arom red brilliant light in the proximal tail of sperm with a diffuse labelling in the distal tail. C) ERα red fluorescence in the sperm mid-piece, together with a faint labelling in the tail. D) ERβ green intense light in the sperm acrosomal region. Inserts: immunonegative absorption controls. Scale bars: 5 μm.

From: Rago V, Aquila S, Panza R, Carpino A. Cytochrome P450arom, androgen and estrogen receptors in pig sperm. Reprod Biol Endocrinol. 2007 Jun 6;5:23.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detection of aromatase in sheep tissues by immunohistochemistry on paraffin sections
Image caption:
a and b) Low- (a) and higher- (b) power sections of the fetal end of a placentome in G1 stained with the prolactin antiserum and showing the intensely stained binucleate cells scattered along the trophoblast, but with a tendency to be more densely accumulated towards the fetal end (scale bars a=200?μm; b=150?μm). (c) High-power section of the trophoblast–endometrium interface in a placentome in G2 stained with the prolactin antiserum. Note the patchy staining of the thin, elongated endometrial epithelial cells (arrowed) in close contact with the intensely stained binucleate trophoblast cells (scale bar=40?μm). (d) Low-power section of the intercotyledonary allantochorion stained with the prolactin antiserum and showing the relatively dense accumulation of binucleate trophoblast cells (arrowed) which remain unstained in this region of the placenta (scale bar=100?μm). (e) High-power section at the edge of a placentome of G1 showing positive staining of the uninucleate trophoblast by the 3β-HSD antiserum. The binucleate cells (arrowed) remain unstained (scale bar=40?μm). (f) Section of the distended endometrial glands at the lateral border of a placentome in G2 stained with 3β-HSD. The basal portions of the endometrial cells lining the glands stain strongly as does the secretory material adhered to the luminal surface of the cells and the accumulated coagulum, possibly as a result of ‘stickiness’ of the material (scale bar=150?μm). (g) High-power section of the intercotyledonary region of the allantochorion in G1 showing positive staining of the cytoplasm of the uninucleate, but not binucleate trophoblast by the 17,20 lyase antiserum (scale bar=40?μm). (h) Section at the fetal end of a placentome from G2 remaining unstained by the aromatase antiserum (scale bar=150?μm). (i) Section of near-term sheep placenta showing strong, positive staining of the uninucleate, but not binucleate, trophoblast by the aromatase antiserum (scale bar=150?μm). (j) High-power section at the fetal end of a placentome in G1 showing strong positive staining of the uninucleate trophoblast by the PR antiserum. The binucleate cells (arrowed) remain unstained (scale bar=40?µm).

From: Wilsher S, Stansfield F, Greenwood RE, Trethowan PD, Anderson RA, Wooding FB, Allen WR. Ovarian and placental morphology and endocrine functions in the pregnant giraffe (Giraffa camelopardalis).Reproduction. 2013 May 21;145(6):541-54.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detection of aromatase in rat prostate by immunohistochemistry on paraffin embedded sections
Image caption:
Immunohistochemical staining of aromatase in prostate of control and BPA-treated rats at doses of 25, 50, 300, or 600 μg/Kg/d for 4 days.

From: Castro B, Sánchez P, Torres JM, Preda O, del Moral RG, Ortega E.
Bisphenol A exposure during adulthood alters expression of aromatase and 5α-reductase isozymes in rat prostate.
PLoS One. 2013;8(2):e55905.

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Mouse anti Human cytochrome p450 aromatase antibody, clone H4 used for the detection of aromatase in human lung tumors by immunohistochemistry on formalin fixed paraffin embedded tissue.
Image caption:
Representative immunohistochemistry of lung tumors for ERα, ERβ, aromatase, EGFR and PR.

From: Stabile LP, Dacic S, Land SR, Lenzner DE, Dhir R, Acquafondata M, Landreneau RJ, Grandis JR, Siegfried JM.
Combined analysis of estrogen receptor beta-1 and progesterone receptor expression identifies lung cancer patients with poor outcome. Clin Cancer Res. 2011 Jan 1;17(1):154-64.

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Mouse anti Human cytochrome p450 aromatase used for the detection of rat aromatase protein by western blotting.
Image caption:
1,25D3 attenuated testosterone-induced aromatase expression but improved 17β-estradiol secretions. Ovarian granulosa cells from rats were pretreated with 1,25D3 (0.1 μM) for 15 min and subsequently treated with 0.1 μg/mL or 1 μg/mL testosterone (denoted by T) or vehicle (denoted by -) for 24 h. (A) Cells were lysed for protein extraction, and cell lysates were subjected to western blot analysis for aromatase protein detection. The level of aromatase (denoted by Arom) expression was estimated by densitometric analyses after normalization with the β-actin signal (n?=?5–6 in each group). (B and C) 17β-Estradiol secretions from the granulosa cells were collected from the medium and assessed over the course of a 24-h period of drug treatments. 17β-Estradiol release data were expressed as the fold-change relative to the vehicle-treated control (n?=?4–5 in each group). The values represent the means?±?SEM. **, p?<?0.005 for comparison with the vehicle-treated group; ##, p?<?0.005 for comparison of testosterone/1,25D3 and testosterone alone; a, p?<?0.05 for comparison of 1,25D3 alone and vehicle-treated group. The values listed within column indicate the means.

From: Lee CT, Wang JY, Chou KY, Hsu MI.
1,25-dihydroxyvitamin D3 increases testosterone-induced 17beta-estradiol secretion and reverses testosterone-reduced connexin 43 in rat granulosa cells.
Reprod Biol Endocrinol. 2014 Sep 20;12:90.

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Mouse anti Human cytochrome p450 aromatase used for the detection of rat aromatase protein by immunoprecipitation and western blotting.
Image caption:
1,25D3 enhanced testosterone-induced aromatase tyrosine phosphorylation and 17β-estradiol secretion during 18–24 h. (A) Rat ovarian granulosa cells were treated with vehicle, testosterone, testosterone plus 1,25D3 or 1,25D3 alone for 24 h (n = 4–5 in each group). Aromatase was immunoprecipitated before being subjected to SDS-PAGE for resolution. The resultant blot was probed using both phosphotyrosine and aromatase antibodies. The expression of aromatase tyrosine phosphorylation (denoted by pTyr-Arom) was normalized to the corresponding aromatase expression and is represented as a percentage relative to the vehicle-treated control. (B) The left panel is a positive control that was both immunoprecipitated and immunoblotted with an anti-aromatase antibody. The lysate was also immunoprecipitated with the anti-aromatase antibody or IgG and immunoblotted with an anti-phosphotyrosine antibody, as shown in the right panel. A weak band in the IgG lane is as a negative control. (C) Cells were treated with testosterone alone or testosterone plus 1,25D3 as indicated for various time periods (n = 4–5 in each group). Each pTyr-Arom level was normalized using the corresponding aromatase level and is represented as a percentage relative to the vehicle-treated control. (D) After treatment for various time periods as indicated, the 17β-estradiol production levels were measured by radioimmunoassay. (E) After treatment with testosterone alone or testosterone plus 1,25D3 for 18 h, the culture medium was removed and then replaced with fresh medium. Cells were continuously treated for another 6 h (18–24 h span). After the final 6 h, medium was collected and the 17β-estradiol secretion levels were measured by radioimmunoassay (n = 6 in each group). The values represent the means ± SEM. #, p < 0.05; ##, p < 0.005 for comparison of the two treatment groups. The values listed within column indicate the means.

From: Lee CT, Wang JY, Chou KY, Hsu MI.
1,25-dihydroxyvitamin D3 increases testosterone-induced 17beta-estradiol secretion and reverses testosterone-reduced connexin 43 in rat granulosa cells.
Reprod Biol Endocrinol. 2014 Sep 20;12:90.