BRD4 antibody (pAb)

RRID: AB_2615059
Aliases: CAP, MCAP, HUNK1, HUNKI
Catalog No: 39909 Format: 100 µl $445 Buy
Catalog No: 39010 Format: 50 µl $260 Buy
Catalog No: 39910 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
180 kDa
Reactivity:
Human, Mouse

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunohistochemistry Validated

Application Notes


Validated Applications:
ChIP: 5 - 10 µl per ChIP
WB: 1:500 - 1:2,000 dilution
IHC (FFPE): 1:1000 dilution

Published Applications:
ChIP-Seq
ChIP

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet; therefore, we recommend a High Salt & Sonication Protocol when preparing nuclear extracts. Visit www.activemotif.com to download the protocol. The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer can be used to aid in detection by Western blot. Individual optimization may be required.

Immunogen

This BRD4 antibody was raised against a peptide within the C-terminal region of human BRD4.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

BRD4 antibody (pAb) tested by ChIP.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 10 µg of chromatin from TNF-α treated HT-29 colorectal adenocarcinoma cells and 5 µl BRD4 antibody. ChIP DNA was used in qPCR with the control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

BRD4 antibody (pAb) tested by Western blot.
Nuclear extract of P19 cells (25 µg / lane) was probed with BRD4 antibody (pAb) at a dilution of 1:1000.

BRD4 antibody (pAb) tested by Immunohistochemistry
Nuclear staining pattern is detected in Formalin-fixed, paraffin-embedded tissue sections from human placenta. Top Panel: BRD4 antibody at 1:1000 dilution. Bottom Panel: No primary antibody (2nd step antibody alone)

Background

BRD4 (Bromodomain-containing protein 4, aka HUNK1) is a chromatin-associated protein with two bromo domains that binds acetylated histone H3 and H4. BRD4 is homologous to Xenopus MCAP, and like MCAP, BRD4 associates with chromosomes during mitosis, predominantly in the non-centromeric regions. A chromosomal aberration involving BRD4 (producing a BRD4-NUT fusion protein) is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. BRD4 is involved in the expression of extracellular matrix gene expression, and its misregulation may be involved in breast cancer progression.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for BRD4 antibody.

qPCR Primers

Active Motif's Human Negative Control Primer Set 1 (Cat. No. 71001) has been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with BRD4 antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting