ANTI-MED28 CLONE 7E1

General description

Mediator of RNA polymerase II transcription subunit 28 (UniProt Q9H204; also known as Endothelial-derived protein 1, Magicin, Mediator complex subunit 28, Merlin and Grb2-interacting cytoskeletal protein, Tumor angiogenesis marker EG-1) is encoded by the MED28 (also known as EG1, FKSG20) gene (Gene ID 80306) in human. MED28 (Magicin) is a subunit of the Mediator complex that is composed of at least 30 subunits and plays an essential role in transcriptional regulation by bridging RNA polymerase II transcriptional machinery and DNA-bound regulatory factors. MED28 is reported to mediate cell surface receptor-mediated signaling and associate with Src-family kinases (e.g. Fyn, Src and Lck), as well as with the Neurofibromatosis 2 (NF2) tumor suppressor merlin. MED28 is associated with both the cytoskeleton and the nucleus, indicating a role as an adaptor/scaffolding protein to relay cellular signals to the cytoskeleton and the nucleus. Homozygous Med28-deletion causes peri-implantation lethality with a loss of pluripotency within the inner cell mass (ICM) of the blastocyst. In addition, Med28-/- embryonic fibroblasts (MEFs) lose the capability to reprogram and Med28 overexpression augments reprogramming efficiency. Upregulated MED28 expression is associated with poor outcome in breast cancer patients, and targeted MED28 downregulation decreases breast cancer cell proliferation and tumor growth.

Specificity

Clone 7E1 detected MED28 in wild-type, but not Med28-/- murine cells. Clone 7E1 detected Cre expression-induced MED28 knockdown in MEFs with floxed Med28 alleles (Li, L., et al. (2015). PLoS One. 10(10):e0140192).

Immunogen

GST-tagged recombinant human MED28 lacking N-terminal 12 amino acids.

Application

Detect MED28 using this mouse monoclonal Anti-MED28 Antibody, clone 7E1, Cat. No. MABC1143, validated for use in Electron Microscopy, Immunocytochemistry, and Western Blotting.

Research Category
Apoptosis & Cancer

Western Blotting Analysis: 1 µg/mL from a representative lot detected MED28 in 10 µg of human embryonic kidney HEK293 cell lysates.

Electron Microscopy Analysis: A representative lot detected MED28 (Magicin) immunoreactivity on cortical actin filaments, colocalizing with that of merlin by immunogold staining of 1% glutaraldehyde-fixed, 0.1% SDS-treated CAD mouse neuronal cells (Wiederhold, T., et al. (2004). Oncogene. 23(54):8815-8825).

Immunocytochemistry Analysis: A representative lot immunostained 4% paraformaldehyde-fixed, 0.25% Triton X-100-permeabilized wild-type, but not Med28-/-, mouse blastocysts, as well as wild-type MEF-dervied iPSC colonies (Li, L., et al. (2015). PLoS One. 10(10):e0140192).

Immunocytochemistry Analysis: A representative lot detected MED28 (Magicin) immunoreactivity colocalized with that of phalloidin-stained actin filament by flourescent immunocytochemistry staining of 0.1% SDS-treated CAD mouse neuronal cells (Wiederhold, T., et al. (2004). Oncogene. 23(54):8815-8825).

Western Blotting Analysis: A representative lot detected a higher MED28 expression in mouse embryonic stem cells (ESCs) than in embryonic fibroblasts (MEFs). Clone 7E1 detected Cre expression-induced MED28 knockdown in MEFs with floxed Med28 alleles (Li, L., et al. (2015). PLoS One. 10(10):e0140192).

Western Blotting Analysis: A representative lot detected ~24 kDa MED28 (Magicin) in human (F293, Gli238, HEK293T, HeLa, MeT5A, T-47D), monkey (COS-7), and mouse (CAD) cell lysates, as well as in Merlin immunoprecipitates from T-47D cytosolic/heavy membrane fractions (Cyt/HMF) and whole cell lysate (Wiederhold, T., et al. (2004). Oncogene. 23(54):8815-8825).

Quality

Evaluated by Western Blotting in RAW267.7 murine macrophage lysate.

Western Blotting Analysis: 1 µg/mL of this antibody detected MED28 in 10 µg of RAW267.7 murine macrophage lysate.

Target description

~20 kDa observed. 19.52 kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.