BRD4 antibody (pAb)

RRID: AB_2615059
Aliases: CAP, MCAP, HUNK1, HUNKI
Catalog No: 39909 Format: 100 µl Discontinued
Catalog No: 39010 Format: 50 µl Discontinued
Catalog No: 39910 Format: 10 µl Discontinued
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

This product has been discontinued:
Please refer to our alternative product Catalog No. 91301

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

*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 for Western blot. For BRD4, we also offer AbFlex® BRD4 Recombinant Antibody (rAb). For details, see Catalog No. 91301.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-qPCR, ChIP-Seq
IF
WB

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

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.

 
BRD4 antibody (pAb) tested by ChIP.

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.

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

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. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • 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;
  • NEU = Neutralizing;
  • WB = Western Blotting