Component of the Mediator complex, a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. This subunit may specifically regulate transcription of targets of the Wnt signaling pathway and SHH signaling pathway.
Defects in MED12 are the cause of Opitz-Kaveggia syndrome (OKS) [MIM:305450]; also known as FG syndrome type 1 (FGS1) or FG syndrome (FGS). OKS is an X-linked disorder characterized by mental retardation, relative macrocephaly, hypotonia and constipation. Defects in MED12 are the cause of Lujan-Fryns syndrome (LUJFRYS) [MIM:309520]; also known as X-linked mental retardation with marfanoid habitus. Clinically, Lujan-Fryns syndrome can be distinguished from Opitz-Kaveggia syndrome by tall stature, hypernasal voice, hyperextensible digits and high nasal root.
Belongs to the Mediator complex subunit 12 family.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling MED12 with ab70842 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - MED12 antibody (ab70842)
All lanes : Anti-MED12 antibody (ab70842) at 0.1 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg Lane 5 : Whole cell lysate from NIH 3T3 cells at 50 µg
Detection of Human MED12 by Immunoprecipitation using Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab70842 for IP at 3 µg/mg lysate. Subsequent WB detection was performed using ab70842 at 1 µg/ml.
ICC/IF image of ab70842 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70842, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Wang L et al. MED12 methylation by CARM1 sensitizes human breast cancer cells to chemotherapy drugs. Sci Adv1:e1500463 (2015).
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Tsutsui T et al. Mediator complex recruits epigenetic regulators via its two cyclin-dependent kinase subunits to repress transcription of immune response genes. J Biol Chem288:20955-65 (2013).
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