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Ab124776

Manufactured by Abcam
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Sourced in United Kingdom, United States

Ab124776 is a laboratory product manufactured by Abcam. It is a core piece of equipment used in scientific research and analysis. The detailed specifications and intended use of this product are not available in this response.

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Lab products found in correlation

Ab22595, by Abcam (1 mentions) Ab27671, by Abcam (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Mda mb 231 htb 26, by American Type Culture Collection (1 mentions) Mcf12f, by American Type Culture Collection (1 mentions) T47d htb 133, by American Type Culture Collection (1 mentions) Nuserum, by Thermo Fisher Scientific (1 mentions) β actin sc 47778, by Santa Cruz Biotechnology (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Ab5593, by Abcam (1 mentions) Ab75200, by Abcam (1 mentions) Sc 365645, by Santa Cruz Biotechnology (1 mentions) Radioimmunoprecipitation assay (ripa), by Applygen (1 mentions) Bca protein assay kit, by CWBIO (1 mentions)

4 protocols using ab124776

1

Western Blot Analysis of Protein Expression

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Protein samples were extracted from cells. Antibodies used include: aromatase (ab124776; Abcam; Cambridge, United Kingdom), V5-tag (ab27671; Abcam), Calnexin (ab22595; Abcam), Gapdh and β-actin (SC-47724 and −47778; Santa Cruz Biotechnology; Dallas, TX, USA). Proteins transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore) were blotted with specific primary antibodies and horseradish peroxidase-conjugated secondary antibodies. Immunoblots were visualized by enhanced chemiluminescence on premium X-ray films (Phenix Research; Candler, NC, USA). Densitometric analyses of images were performed using Image Processing and Analysis in Java (ImageJ) software from three independent experiments.
Molehin D., Castro-Piedras I., Sharma M., Sennoune S.R., Arena D., Manna P.R, & Pruitt K. (2018). Aromatase Acetylation Patterns and Altered Activity in Response to Sirtuin Inhibition. Molecular cancer research : MCR, 16(10), 1530-1542.
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2

Breast Cancer and Adrenocortical Cell Lines

Cited 2 times
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Human normal mammary epithelial MCF10A (CRL-10317) and MCF12F (CRL-10783) cells, and ER+ MCF7 (HTB-22), MDA-MB-361 (HTB-27), and T-47D (HTB-133), and triple negative MDA-MB-468 (HTB-132), BT-549 (HTB-19), and MDA-MB-231 (HTB-26) breast cancer cells were purchased from ATCC (Manassas, VA), and were maintained in specific growth media containing antibiotics [15 (link)], according to instructions from the ATCC. Human adrenocortical tumor H295R (CRL-2128) cell line was obtained from ATCC and were cultured in DMEM/F12 with 1% ITS plus 2.5% NuSerum containing penicillin/streptomycin (Invitrogen, Carlsbad, CA) [16 (link)].
LBH589 (Panobinostat), MS-275 (Entinostat), and SB939 (Practinostat) were purchased from APExBIO (Houston, TX), and Vorinostat (SAHA), Sirtuin (SIRT) 1/2 inhibitors IV and VII, and romidepsin (Istodax, FR228) were purchased from Millipore-Sigma (St. Louis, MO). StAR (Ab133657 or Ab180804; AbCam, Cambridge, MA), aromatase (Ab124776, AbCam), and acetyl lysine (05-515, AbCam), and β-actin (sc47778, Santa Cruz Biotechnology) antibodies (Abs) were purchased from the indicated commercial sources.
Manna P.R., Ahmed A.U., Vartak D., Molehin D, & Pruitt K. (2018). Overexpression of the steroidogenic acute regulatory protein in breast cancer: Regulation by histone deacetylase inhibition. Biochemical and biophysical research communications, 509(2), 476-482.
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3

Protein Expression Analysis in Granulosa Cells

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Cellular proteins were extracted from ovaries and cultured granulosa cells with RIPA (Applygen Technologies Inc., Beijing, China) supplemented with 1 mM PMSF serine protease inhibitor. The protein content was measured using a BCA Protein Assay Kit according to the manufacturer’s guidelines (CWBIO, Beijing, China). Western blotting was conducted as described in a previous report [24 (link)]. A total of 20 micrograms of protein was electrophoresed on a 12% SDS-polyacrylamide gel and transferred to PVDF membranes. The levels of the Bmal1 and other proteins involved in Lepr function and E2 synthesis were measured using specific antibodies. Antibodies against Bmal1 (1/500; sc-365645, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), Lepr (1/500; ab5593, Abcam, Inc., Cambridge, MA, USA), Fshr (1/1000; ab75200, Abcam, Inc.), Cyp19a1 (1/1000; ab124776, Abcam, Inc.), and Cyp11a1 (1/500; #14217, CST, Inc., Danvers, MA, USA) were used to detect the levels of these proteins in granulosa cells.
Chu G., Ma G., Sun J., Zhu Y., Xiang A., Yang G, & Sun S. (2019). Leptin Receptor Mediates Bmal1 Regulation of Estrogen Synthesis in Granulosa Cells. Animals : an Open Access Journal from MDPI, 9(11), 899.
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4

Aromatase Acetylation Sites in MCF-7 Cells

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MCF-7 cells were seeded and incubated at 37°C with either 20% or 2.5% O2. Cells were treated when they reached 70–80% confluence with either DMSO or 32nM inhibitor IV for 10mins. Cells were lysed with acKIP buffer and protein extracts incubated with 4μg anti-aromatase antibody (ab124776; Abcam) overnight. Protein-G dynabeads were added to the antibody-protein-complex and incubated for 2 hours. Immunoprecipitates were shipped to Applied Biomics Inc. (Hayward, CA, USA) for acetylation site identification by LC-MS/MS mass spectrometry.
Molehin D., Castro-Piedras I., Sharma M., Sennoune S.R., Arena D., Manna P.R, & Pruitt K. (2018). Aromatase Acetylation Patterns and Altered Activity in Response to Sirtuin Inhibition. Molecular cancer research : MCR, 16(10), 1530-1542.
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