Application

Detect BRD4 Serine 492 phosphorylation using this rabbit polyclonal Anti-phospho BRD4 (Ser492), Cat. No. ABE1451, validated for use in Immunocytochemistry, Dot Blot and Western Blotting.

Research CategoryEpigenetics & Nuclear Function

Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected nuclear Brd4 pSer493 (equivalent to human Ser492) immunoreactivity in 12-day cultured E16.5 embryonic mouse cortical neurons (Courtesy of Erica Korb, Ph.D. Rockefeller University, New York, U.S.A.).University, New York, U.S.A.).

Western Blotting Analysis: A 1:500 dilution from a representative lot detected Brd4 Ser493 (equivalent to human Ser492) phosphorylation in lysate from 12-day cultured E16.5 embryonic mouse cortical neurons. Prior phosphatase treatment of the lysate abolished target band detection (Courtesy of Erica Korb, Ph.D. Rockefeller University, New York, U.S.A.).

Dot Blot Analysis: A representative lot detected BRD4 phosphopeptide with pSer492 or dual pSer492/494 phosphorylation (equivalent to mouse Ser493/495), but not the corresponding non-phosphorylated peptide or peptide with only pSer494 phosphorylation (Korb, E., et al. (2015). Nat. Neurosci. 18(10):1464-1473).

Western Blotting Analysis: A representative lot detected Brd4 Ser493 (equivalent to human Ser492) phosphorylation induction upon BDNF stimulation of cultured E16.5 embryonic mouse cortical neurons from C57BL/6 mice. Pre-treatment of neurons with CK2 inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB; Cat. No. 218697) before BDNF stimulation or phosphatase treatment of the lysate prior to immunoblotting abolished target band detection (Korb, E., et al. (2015). Nat. Neurosci. 18(10):1464-1473).

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.

General description

Bromodomain-containing protein 4 (UniProt O60885; also known as MCAP, Mitotic chromosome-associated protein, Protein HUNK1) is encoded by the BRD4 (also known as HUNK1, MCAP) gene (Gene ID 23476) in human. BRD4 is a BET (bromodomain and extra-terminal) protein family member that functions as a chromatin reader by binding acetylated lysines in histones. BRD4 is an important transcription regulator in many cell types, including transcription in response to external cues. BRD4 plays an essential role in neuronal function and mediates the transcriptional regulation underlying learning and memory. BRD4 regulates IEG transcription in neurons upon phosphorylation by casein kinase 2 (CK2), which is activated in response to neuronal stimulation. Brd4 knockout in mice is lethal, while Brd4 function blockage by inhibitor JQ1 (Cat. No. 500586) treatment is reported to affect the transcription of critical synaptic proteins, resulting in memory deficits and decreased seizure susceptibility in mice.

Immunogen

KLH-conjugated linear peptide corresponding to a human BRD4 sequence containing phosphorylated Ser492 (equivalent to mouse Ser493).

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Affinity purified.

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide.

Format: Purified

Quality

Evaluated by Western Blotting in human iPSC lysate.

Western Blotting Analysis: A 1:1000 dilution of this antibody detected Brd4 Ser492 phosphorylation in 10 µg of human induced pluripotent stem cell (iPSC) lysate.

Specificity

The target phospho-serine residue is present in all human (pSer492) and murine (pSer493) BRD4 spliced isoforms reported by UniProt (O60885 and Q9ESU6). This polyclonal antibody is shown to exhibit stronger immunoreactivity toward BRD4 phosphopeptide with dual pSer492/494 (equivalent to mouse pSer493/495) phosphorylation than the corresponding peptide with only pSer492 (equivalent to mouse Ser493) phosphorylation (Korb, E., et al. (2015). Nat. Neurosci. 18(10):1464-1473).

Storage and Stability

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

Target description

~140/200 kDa observed. 152.2/155.9 kDa (human/mouse isoform Long), 80.46/80.61 kDa (human/mouse isoform Short), 88.29 kDa (human isoform B) calculated. Uncharacterized bands may be observed in some lysate(s).